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Blood, 1 August 2008, Vol. 112, No. 3, pp. 836-839.
Prepublished online as a Blood First Edition Paper on May 22, 2008; DOI 10.1182/blood-2007-12-126979.
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NEOPLASIA
Brief Report
Restrictive usage of monoclonal immunoglobulin  light chain germline in POEMS syndrome
Daijiro Abe1,
Chiaki Nakaseko1,
Masahiro Takeuchi1,
Hiroaki Tanaka1,
Chikako Ohwada1,
Emiko Sakaida1,
Yusuke Takeda1,
Kayo Oda1,
Shinichi Ozawa1,
Naomi Shimizu1,
Shinichi Masuda1,
Ryuko Cho1,
Miki Nishimura1,
Sonoko Misawa2,
Satoshi Kuwabara2, and
Yasushi Saito1
1 Division of Hematology, Department of Clinical Cell Biology, and
2 Department of Neurology, Chiba University Graduate School of Medicine, Chiba, Japan
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Abstract
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POEMS syndrome is a rare plasma cell disorder characterized by peripheral neuropathy, monoclonal gammopathy, and high levels of serum vascular endothelial growth factor, the pathogenesis of which remains unclear. A unique feature of this syndrome is that the proliferating monoclonal plasma cells are essentially  -restricted. Here we determined complete nucleotide sequences of monoclonal immunoglobulin light chain (IGL) variable regions in 11 patients with POEMS syndrome. The V-region of the Ig gene of all 11 patients was restricted to the V1 subfamily. Searching for homologies with IGL germlines revealed that 2 germlines, IGLV1-44*01 (9/11) and IGLV1-40*01 (2/10), were identified, with an average homology of 91.1%. The IGLJ3*02 gene was used in 11 of 11 re-arrangements with an average homology of 92.2%. These data suggest that the highly restricted use of IGL V1 germlines plays an important role in the pathogenesis of POEMS syndrome.
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Introduction
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POEMS syndrome is a rare plasma cell disorder characterized by peripheral neuropathy, monoclonal gammopathy, and other paraneoplastic features, including organomegaly, endocrinopathy, skin changes, edema, and effusions.1–3 The pathogenesis of this disease is not well understood, but overproduction of vascular endothelial growth factor (VEGF), presumably secreted by plasmacytoma cells or platelets, is considered to be responsible for the characteristic symptoms.4–6 Treatment for POEMS syndrome includes surgical resection or radiation for plasmacytoma, and chemotherapy, especially high-dose melphalan, followed by autologous peripheral blood stem cell transplantation.7–9 Recently, thalidomide,10,11 lenalidomide,12 and anti-VEGF monoclonal antibodies have also been used.13–15
A unique feature of this syndrome is that the proliferating monoclonal plasma cells are essentially -restricted in most cases, and  type is extremely rare.16 However, the pathogenetic role of -type M protein in POEMS syndrome is unclear. Here we determined complete nucleotide sequences of monoclonal immunoglobulin light chain (IGL) variable regions in patients with POEMS syndrome and found that V germline usage is highly restricted to the V1 subfamily and very limited germlines.
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Methods
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Study design
This study was approved by the Institutional Review Board of Chiba University Hospital.
Patients
A total of 15 patients (9 men and 6 women; median age at diagnosis, 54.7 years) were included in the study (Table 1). All patients met the criteria for the diagnosis of POEMS syndrome17 and had the 5 distinctive features of POEMS. The serum monoclonal immunoglobulin light chain was -type in all patients. The serum monoclonal heavy chain component was IgG in 7 patients and IgA in 8 patients. Informed consent was obtained from the patients in accordance with the Declaration of Helsinki.
Amplification of monoclonal chain gene and sequencing analysis
Total RNA was extracted from bone marrow mononuclear cells of patients with POEMS syndrome using TRIzol (Invitrogen, Carlsbad, CA), and single-stranded cDNA was synthesized using Superscript II (Invitrogen). The V-J region of the IGL gene was amplified by reverse-transcribed polymerase chain reaction (PCR) using 5' degenerate primers for the V1-V2-V3 consensus leader lesion (5'-ATGGCCKGSWYYSYTCTCCTC-3') and 3' primers matching the consensus upstream part of the c exon (5'-CTCCCGGGTAGAGAAGTCACT-3') as previously described.18 To identify the amount of monoclonal PCR products, we used heteroduplex analysis in which PCR products are denatured at 95°C for 5 minutes and subsequently renatured at 4°C for 1 hour to induce homoduplex or heteroduplex formation.19 When sufficient monoclonal PCR product was available for direct sequencing, PCR products were subjected to cycle sequencing using the aforementioned primers. If the amount was small, homoduplex bands on the polyacrylamide gel after heteroduplex analysis were excised and monoclonal PCR fragments were eluted from the solubilized gel slice using QIAEX II (Qiagen, Valencia, CA) and subjected to cycle sequencing.
Sequence data were analyzed using the database of the International ImMunoGeneTics information system (Montpellier, France), and mutations were identified by comparison with the germline sequence. The average mutation rate of the IGL gene was calculated as pre-viously described.20
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Results and discussion
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Of 15 patients with POEMS syndrome who had -type M protein, nucleotide sequencing of the IGL gene was successful in 11 (Table 1). The remaining 4 patients' samples had none or too little of the monoclonal band to sequence in heteroduplex analysis. Sequence analysis revealed that the V-region of the monoclonal Ig gene of all 11 patients was restricted to the V1 subfamily. Searching for homologies with IGL germlines revealed that they were derived exclusively from IGLV1-44*01 (9 patients) or IGLV1-40*01 (2 patients). Complete nucleotide and amino acid sequences are shown in Figure 1. IGLV rearrangements were mutated with homologies to germline ranging from 81.5% to 96.1% (average homology, 91.1%). In the IGL-J region, the IGLJ3*2 gene was found in 11 of 11 patients. The rearrangements were mutated with homologies to germline ranging from 84.1% to 100% (average homology, 92.2%). All complementarity-determining region 3 (CDR3) were composed of 11 amino acids, with identical acidic isoelectric point values of 13.0 and similar molecular weights ranging from 1441.6.7 to 1552.8 (median, 1524.5). The average nucleotide mutation rate was 8.6% with 7.5% in the framework regions (FWRs) and 11.8% in CDRs. The ratio of replacement/silent (R/S) mutations was higher in the CDRs than in the FWRs.
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Figure 1. Complete nucleotide and amino acid sequences of monoclonal IGL gene in POEMS syndrome. (A) Complete nucleotide sequences of 2 cases (nos. 1 and 2) and previously reported 2 cases by Soubrier et al (A and B)18 with the IGLV1-40*01 germline. (B) Complete nucleotide sequences of 9 cases with the IGLV1-44*01 germline. (C) Complete amino acid sequences of these 13 cases. FR indicates framework; CDR, complementarity-determining region; hyphens represent nucleotide or amino acid identity; dots represent gaps.
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In this study, the monoclonal IGL gene could not be analyzed in 4 patients' samples. Two cases had very small amounts of monoclonal band, and the other 2 had no visible monoclonal band at amplification by V1-3 degenerate primers. The latter 2 cases might have monoclonal IGL genes belonging to a V subfamily other than V1-3.
Analyses of monoclonal light chain genes have been extensively carried out in plasma cell dyscrasias by others. González et al reviewed 4 reports of IGL gene usage in Ig- type multiple myeloma and showed that mainly the V1 (27%), V2 (28%), and V3 (41%) subfamilies were used to a similar extent in -type myeloma and that V4 through V10 subfamily usage was extremely rare (4%).21 Within these families, no clear preference for individual gene segments was observed. Comenzo et al reported the results of IGL germline gene analysis in 39 patients with AL amyloidosis.22 The clonal AL gene subfamily belonged to different subfamilies, including V1 (26%), V2 (13%), V3 (15%), V6 (28%), and V1 (18%). Other germline genes were used, of which the IGLV6S1 germline of the V6 subfamily was relatively dominant. We found that the monoclonal IGLV gene of all 11 POEMS patients belonged to the V1 subfamily with only 2 germlines of V1 represented. There has been only one other report of IGLV gene analysis in POEMS syndrome by a French group.18 In that study, Soubrier et al18 found that both of the 2 cases analyzed were derived from the V1-IGLV1-40 germline, the same as in our study (Figure 1A,C). Taken together with our results, currently available data on 13 cases indicate that IGLV genes are restricted to the 2 V1 germlines: IGLV1-44 in 9 and IGLV1-40 in 4 across 2 ethnically different populations.
Why is the light chain germline repertoire so restrictive and what links exist between germline gene use and VEGF in POEMS syndrome? Perfetti et al reported that amyloid V regions were highly mutated in AL amyloidosis and amyloidogenic light chains undergo antigen-driven selection.23 Because the average nucleotide mutation rate and the R/S mutation ratio were higher in the CDRs than in the FWRs, restricted light chains might undergo antigen-driven selection in POEMS. These restricted chains might interact with VEGF-related protein and play a role in VEGF secretion. In myeloma, interactions between light-chain CDR3 sites and proteins, such as Tamm-Horsfall proteins, have been previously described.24 However, the higher R/S mutation ratio in the CDRs and retention of cysteine residues in FR1 and FR3, which were critical to structure, suggest that these specific light chains might interact with VEGF-related protein at FRWs but not CDRs like B-cell superantigen.25
In conclusion, IGL-M protein in POEMS syndrome belongs to the V1 subfamily and is markedly restricted to a very limited number of germlines. Although exact mechanisms responsible for this syndrome are still unclear, these data suggest that the restricted use of IGL plays an important role in pathogenesis and may represent a new clinical entity. Further studies are required to clarify the relationship between high VEGF levels and restricted usage of V genes in POEMS syndrome.
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Authorship
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Contribution: D.A., C.N., M.T., H.T., C.O., and Y.S. designed the research; D.A. and C.N. performed research, analyzed data, and wrote the paper; E.S., Y.T., K.O., S.O., N.S., S. Masuda, R.C., and M.N. designed the research and analyzed data; S. Misawa, and S.K. organized collection of patient samples; and all authors approved the final version of the paper.
Conflict-of-interest disclosure: The authors declare no competing financial interests.
Correspondence: Chiaki Nakaseko, Division of Hematology, Department of Clinical Cell Biology, Chiba University Graduate School of Medicine, 1-8-1, Inohana, Chuo-ku, Chiba City, Chiba, 260-8670, Japan; e-mail: chiaki-nakaseko{at}faculty.chiba-u.jp.
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Acknowledgments
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The authors thank Ms Megumi Naito for technical assistance, Ms Kimie Shibuya for secretarial assistance, and Dr Takeshi Tokuhisa and Dr Akemi Sakamoto of the Department of Developmental Genetics, Chiba University Graduate School of Medicine, for helpful discussions.
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Footnotes
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Submitted December 1, 2007;
accepted April 19, 2008.
Prepublished online as Blood First Edition Paper, May 22, 2008
DOI: 10.1182/blood-2007-12-126979
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 USC section 1734.
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References
|
|---|
- Bardwick PA, Zvaifler NJ, Gill GN, Newman D, Greenway GD, Resnick DL. Plasma cell dyscrasia with polyneuropathy, organomegaly, endocrinopathy, M protein, and skin changes: the POEMS syndrome. Report on two cases and a review of the literature. Medicine (Baltimore). 1980;59:311–322.[Medline]
[Order article via Infotrieve]
- Dispenzieri A, Kyle RA, Lacy MQ, et al. POEMS syndrome: definitions and long-term outcome. Blood. 2003;101:2496–2506.[Abstract/Free Full Text]
- Miralles GD, O'Fallon JR, Talley NJ. Plasma-cell dyscrasia with polyneuropathy: the spectrum of POEMS syndrome. N Engl J Med. 1992;327:1919–1923.[Abstract]
- Watanabe O, Arimura K, Kitajima I, Osame M, Maruyama I. Greatly raised vascular endothelial growth factor (VEGF) in POEMS syndrome. Lancet. 1996;347:702.[Medline]
[Order article via Infotrieve]
- Watanabe O, Maruyama I, Arimura K, et al. Overproduction of vascular endothelial growth factor/vascular permeability factor is causative in Crow-Fukase (POEMS) syndrome. Muscle Nerve. 1998;21:1390–1397.[CrossRef][Medline]
[Order article via Infotrieve]
- Hashiguchi T, Arimura K, Matsumuro K, et al. Highly concentrated vascular endothelial growth factor in platelets in Crow-Fukase syndrome. Muscle Nerve. 2000;23:1051–1056.[CrossRef][Medline]
[Order article via Infotrieve]
- Kuwabara S, Hattori T, Shimoe Y, Kamitsukasa I. Long term melphalan-prednisolone chemotherapy for POEMS syndrome. J Neurol Neurosurg Psychiatry. 1997;63:385–387.[Abstract/Free Full Text]
- Dispenzieri A, Moreno-Aspitia A, Suarez GA, et al. Peripheral blood stem cell transplantation in 16 patients with POEMS syndrome, and a review of the literature. Blood. 2004;104:3400–3407.[Abstract/Free Full Text]
- Kuwabara S, Misawa S, Kanai K, et al. Autologous peripheral blood stem cell transplantation for POEMS syndrome. Neurology. 2006;66:105–107.[Abstract/Free Full Text]
- Sinisalo M, Hietaharju A, Sauranen J, Wirta O. Thalidomide in POEMS syndrome: case report. Am J Hematol. 2004;76:66–68.[CrossRef][Medline]
[Order article via Infotrieve]
- Kim SY, Lee SA, Ryoo HM, Lee KH, Hyun MS, Bae SH. Thalidomide for POEMS syndrome. Ann Hematol. 2006;85:545–546.[CrossRef][Medline]
[Order article via Infotrieve]
- Dispenzieri A, Klein CJ, Mauermann ML. Lenalidomide therapy in a patient with POEMS syndrome. Blood. 2007;110:1075–1076.[Free Full Text]
- Straume O, Bergheim J, Ernst P. Bevacizumab therapy for POEMS syndrome. Blood. 2006;107:4972–4973; author reply 4973–4974.[Free Full Text]
- Badros A, Porter N, Zimrin A. Bevacizumab therapy for POEMS syndrome. Blood. 2005;106:1135.[Free Full Text]
- Kanai K, Kuwabara S, Misawa S, Hattori T. Failure of treatment with anti-VEGF monoclonal antibody for long-standing POEMS syndrome. Intern Med. 2007;46:311–313.[CrossRef][Medline]
[Order article via Infotrieve]
- Dispenzieri A. POEMS syndrome. Blood Rev. 2007;21:285–299.[CrossRef][Medline]
[Order article via Infotrieve]
- Dispenzieri A. POEMS syndrome. Hematology Am Soc Hematol Educ Program. 2005;360–367.
- Soubrier M, Labauge P, Jouanel P, Viallard JL, Piette JC, Sauvezie B. Restricted use of Vlambda genes in POEMS syndrome. Haematologica. 2004;89:ECR02.[Free Full Text]
- Langerak AW, Szczepanski T, van der Burg M, Wolvers-Tettero IL, van Dongen JJ. Heteroduplex PCR analysis of rearranged T cell receptor genes for clonality assessment in suspect T cell proliferations. Leukemia. 1997;11:2192–2199.[CrossRef][Medline]
[Order article via Infotrieve]
- Chang B, Casali P. The CDR1 sequences of a major proportion of human germline Ig VH genes are inherently susceptible to amino acid replacement. Immunol Today. 1994;15:367–373.[CrossRef][Medline]
[Order article via Infotrieve]
- González D, van der Burg M, Garcia-Sanz R, et al. Immunoglobulin gene rearrangements and the pathogenesis of multiple myeloma. Blood. 2007;110:3112–3121.[Abstract/Free Full Text]
- Comenzo RL, Wally J, Kica G, et al. Clonal immunoglobulin light chain variable region germline gene use in AL amyloidosis: association with dominant amyloid-related organ involvement and survival after stem cell transplantation. Br J Haematol. 1999;106:744–751.[CrossRef][Medline]
[Order article via Infotrieve]
- Perfetti V, Ubbiali P, Vignarelli MC, et al. Evidence that amyloidogenic light chains undergo antigen-driven selection. Blood. 1998;91:2948–2954.[Abstract/Free Full Text]
- Ying WZ, Sanders PW. Mapping the binding domain of immunoglobulin light chains for Tamm-Horsfall protein. Am J Pathol. 2001;158:1859–1866.[Abstract/Free Full Text]
- Levinson AI, Kozlowski L, Zheng Y, Wheatley L. B-cell superantigens: definition and potential impact on the immune response. J Clin Immunol. 1995;15(suppl):26–36.[CrossRef]
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Abstract
Introduction
