Role of platelet-derived growth factor-AB in tumour growth and angiogenesis in relation with other angiogenic cytokines in multiple myeloma

Angiogenesis is a complex process essential for the growth, invasion, and metastasis of various malignant tumours, including multiple myeloma (MM). Various angiogenic cytokines have been implicated in the angiogenic process. Among them, platelet-derived growth factor-AB (PDGF-AB) has been reported to be a potent stimulator of angiogenesis in many solid tumours and haematological malignancies, including MM. The aim of the study was to investigate the relationship between PDGF-AB, microvascular density (MVD), and various angiogenic cytokines, such as basic fibroblast growth factor (b-FGF), angiogenin (ANG), and interleukin-6 (IL-6), in MM patients. Forty-seven MM patients before treatment, 22 of whom were in plateau phase, were studied. We determined the serum levels of the aforementioned cytokines and MVD in bone marrow biopsies before and after treatment. Mean serum values of PDGF-AB, b-FGF, ANG, and MVD were significantly higher in patients compared with controls and with increasing disease stage. Significant positive correlations were observed between serum PDGF-AB, ANG, and IL-6 levels and MVD. Furthermore, we found significant positive correlations between PDGF-AB and b-FGF, IL-6, ANG, and β2 microglobulin. We also found that patients with high MVD had statistically significantly higher serum levels of PDGF-AB when a median MVD value of 7.7 was used as the cutoff point. Furthermore, a significant difference was found in serum levels of PDGF-AB between pre- and post-treatment patients. Finally, survival time was significantly higher in the low MVD group versus the high MVD group (76 vs 51 months). Our results showed that there is a strong positive correlation between PDGF-AB and the studied angiogenic cytokines and MVD. It seems that PDGF-AB plays a role in the complex network of cytokines inducing bone marrow neovascularization in patients with MM.     

Expression of endoglin (CD105) in cervical cancer

In this study, we have investigated the role of endoglin (CD105), a regulator of transforming growth factor (TGF)-β₁ signalling on endothelial cells, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor-A (VEGF-A) in cervical cancer. We have measured the number and determined the location of both newly formed (CD105-positive) and the overall number of (CD31-positive) blood vessels, and bFGF and VEGF-A expression using immunohistochemistry in 30 cervical carcinoma specimens. Vascular endothelial growth factor-A mRNA expression was determined using RNA-in situ hybridisation. CD105- and CD31-positive vessels and bFGF- and VEGF-A-positive cells were predominantly present in the stroma. The presence of CD105- and CD31-positive vessels in the stroma did neither correlate with the number of VEGF-A-positive cells nor the number of bFGF-positive cells. However, the number of CD105- and CD31-positive vessels was associated with the expression of VEGF-A mRNA in the epithelial cell clusters (P=0.013 and P=0.005, respectively). The presence of CD105-positive and CD31-positive vessels was associated with the expression of αvβ6 (a TGF-β₁ activator; P=0.013 and P=0.006, respectively). Clinically, the number of CD105-positive vessels associated with the number of lymph node metastasis (P<0.001). Furthermore, the presence of CD105-positive vessels within the epithelial cell clusters associated with poor disease-free survival (P=0.007).     

The combination of intermediate doses of thalidomide and dexamethasone reduces bone marrow micro-vessel density but not serum levels of angiogenic cytokines in patients with refractory/relapsed multiple myeloma

The aim of the study was the evaluation of anti-angiogenic activity of the combination of intermediate doses of thalidomide and dexamethasone in patients with refractory/relapsed myeloma. Twenty-five patients were included in the study. Microvessel density (MVD) was evaluated in marrow biopsies before and after treatment. Serum levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (b-FGF), tumor necrosis factor-alpha (TNF-alpha), which have angiogenic potential and interleukin-6 (IL-6), IL-1beta, soluble IL-6 receptor (sIL-6R), and transforming growth factor-beta (TGF-beta) which are involved in the disease biology, were measured before treatment and then every 2 weeks for 8 weeks. Pretreatment levels of MVD, VEGF, b-FGF, IL-6, sIL-6R were increased in the patients compared to controls. The overall response rate to therapy was 72%. The administration of the combined regimen produced a significant reduction in MVD in responders. However, an increase in serum levels of VEGF, b-FGF, IL-6, sIL-6R was observed post-treatment in responders. In contrast, serum levels of TNF-alpha, TGF-beta, IL-1beta did not differ between patients and controls and remained unchanged during the study. These results suggest that the combination of thalidomide plus dexamethasone is an effective treatment for myeloma reducing MVD marrow levels but not serum levels of angiogenic cytokines or cytokines implicated in myeloma biology.     

多发性骨髓瘤的治疗

Multiple Myeloma(MM)is characterised by the accumulation of malignanat plasma cells in the bone marrow producing a monocolonal immungoglobulin.The standard conventiona therapy in the combination of melphalan and prednisone resulting in a response rate of 40%-60% and in a median survival time of approximately 3 years.In order to improve the therapeutic efficacy va rious combination regimens have been tested.Most randomized trials have frailed to show a significant improvement in survival time when combination chemotherapy is used instead of melphalan with or without prednisone.The benefit of maintenance therapy with interferon-alpha has been demonstrated.The toxicity of interferon-alpha,which may reduce the quality of life,should be considered.Recently,myeloma-treatment has been modified.High-dose chemotherapy accompaniced by hematopoietic stem-cell support via autologous transplant is recommended up to the age of 65-70 years.First results from a French study comparing single versus double autologous transplantaiton have shown a benefit in terms of event-free survival for the sequential approach.Vaccinations as an adoptive immuntherapy to treat minimal residual dsease are umder way.The mortality rate of allogeneic transplantation of hemaatopoietic stem cells has been reduced in the last 5 years.The use of reduced conditioning regimens or the partial depletion of T cells in peripheral blood stem cell transplants in an effort to decrease transplant related mortality are promising approaches.Thalidomide and its derivates are a new class of agents with independent anti-tumour activity in MM.Encouraging results with this antiangiogenic therapy in phase Ⅱ trials have been reported.Supportive therapies,such as the treatment of anaemia with erythropoietin,the management of renal failure and the use of bisphosphonates,improve the life quality of MM patients.     

Potential relationship and clinical significance of miRNAs and Th17 cytokines in patients with multiple myeloma

We evaluated the potential relationship between miRNAs and Th17 cytokines in multiple myeloma (MM) patients. Twenty-seven newly diagnosed myeloma patients and eight normal donors were studied. We determined that the relative expression levels of miR-15a/16, miR-34a, miR-194 in MM patients were significantly lower than those in the healthy controls with exception for miR-181a/b, which showed significantly higher in MM patients (P < 0.05). In contrast, the levels of IL-17, IL-21 and IL-27 were up-regulated in MM patients compared to healthy controls while IL-22 was down-regulated (P < 0.05). The expression patterns of them were differentially present in various groups according to the International Staging System (ISS) criteria. Up-regulated IL-17, IL-21 and IL-27 may potentially down-regulate the expression of several miRNAs in MM patients. Establishment of the relationship may be useful for understanding the pathogenesis of MM and for clinical diagnosis of the disease.     

Thalidomide: present and future in multiple myeloma

Multiple myeloma continues to be an incurable disease. The understanding of the disease’s pathophysiology has significantly improved over the past few years, partly due to the discovery of the role of immunomodulatory agents and the study of their mechanism of action. Thalidomide, the first of the immunomodulatory family to be used in the management of multiple myeloma, proved not only to be effective in the treatment of multiple myeloma, but also instigated a wide range of in vitro and in vivo studies to define the pathophysiology of the plasma cell dyscrasia. The attention thalidomide has received in the past and recent history has not been without a price. The drug has a side-effect profile that, if managed appropriately, provides the most unique active molecule in the management of the disease, where it maintains the same response rate in newly diagnosed patients as in advanced relapsed/refractory multiple myeloma patients.     

多发性骨髓瘤患者OPN、CD44v6的表达与病情进展的关系

背景与目的:骨桥蛋白(osteopontin,OPN)与其受体--CD44受体和整合素受体结合后,参与骨重建、骨质重吸收、血管重塑、癌细胞浸润和转移等过程.为此本研究检测了多发性骨髓瘤(multiple myeloma,MM)患者的OPN和变异的CD44受体(CD44v6)水平,以探讨其与MM病情进展的关系.方法:32例MM患者,分为A组(包括初发和复发病例)和B组(化疗后病情稳定)2组,同时收集外伤骨折或非肿瘤良性贫血患者15例作为对照.采用酶链免疫吸附试验(ELISA)分别检测了患者和对照组的单个核细胞及骨髓基质细胞(bone marrow mononuclear eells,BMSCs)培养上清液的0PN、CD44v6水平并进行相关分析.结果:A组(19例)患者的0PN表达水平显著高于B组(13例)和对照组患者(P<0.05).对A组中的14例患者和B组中的10例患者进行了CD44v6水平检测,其结果A组的CD44v6表达水平显著高于B组和对照组患者(P<0.05),B组的CD44v6水平显著高于对照组(P<0.05);0PN水平与CD44v6(r=0.52,P=0.000)、恶性浆细胞数(r=0.74,P=0.000)、M蛋白水平(r=0.53,P=0.014)、血β2-MG水平(r=0.62,P=0.002)均呈正相关.结论:0PN和CD44v6水平升高与删的发生和病情进展有关;可能与MM的肿瘤负荷、疾病阶段和肿瘤浸润有关.     

多发性骨髓瘤44例临床分析

目的:探讨多发性骨髓瘤的临床特点。方法:回顾性分析西安交大二院血液科2001年～2006年收治的44例多发性骨髓瘤患者的临床资料。结果:发病中位年龄为58.23岁,首发症状为骨痛(40%),乏力(16.3%)。血清β2-MG,CRP升高者及血清白蛋白水平降低者中位生存期与上述三值正常者相比均有显著性差异。结论:生存分析显示β2微球蛋白和C反应蛋白与疾病预后相关。     

Murine 5T multiple myeloma cells induce angiogenesis in vitro and in vivo

Multiple myeloma is a B cell malignancy. Recently, it has been demonstrated that bone marrow samples of patients with multiple myeloma display an enhanced angiogenesis. The mechanisms involved seem to be multiple and complex. We here demonstrate that the murine 5T multiple myeloma models are able to induce angiogenesis in vitro by using a rat aortic ring assay and in vivo by determining the microvessel density. The rat aortic rings cultured in 5T multiple myeloma conditioned medium exhibit a higher number of longer and more branched microvessels than the rings cultured in control medium. In bone marrow samples from 5T multiple myeloma diseased mice, a statistically significant increase of the microvessel density was observed when compared to bone marrow samples from age-matched controls. The angiogenic phenotype of both 5T multiple myeloma cells could be related, at least in part, to their capacity to produce vascular endothelial growth factor. These data clearly demonstrate that the 5T multiple myeloma models are good models to study angiogenesis in multiple myeloma and will allow to unravel the mechanisms of neovascularisation, as well as to test new putative inhibitors of angiogenesis.     

Upregulation of CD200 is associated with regulatory T cell expansion and disease progression in multiple myeloma

Immune dysfunction is an important feature of multiple myeloma (MM) leading to infections, enhancement of tumour growth and resistance to chemotherapy. The overexpression of CD200, expansion of T regulatory (Treg) cell and increased levels of immune modulatory cytokines like IL10, IL6 and transforming growth factor beta (TGFβ) were suggested to have a role in this context. The aim of this study was to assess CD200 expression, Treg percentage by flow cytometry and immune modulatory cytokines (IL10, IL6, TGFβ) by enzyme‐linked immunosorbent assay in MM patients at diagnosis. This study included 50 MM patients at diagnosis and 20 healthy controls. The positive CD200 expression was detected in 72% of MM patients. Among the CD200 positive group, 4/13 patients (30.8%) were classified as stage I, 18/23 (78.3%) were in stage II and 14/14 (100%) were in stage III; according to International scoring system. Treg percentage was significantly higher in stage III, followed by stage II then stage I (p < 0.01). Serum IL6, IL10 and TGFβ were significantly higher in MM patients as compared with controls (p < 0.01, p < 0.01, p < 0.05, respectively). The increased expression of CD200 and Treg percentages was associated with increased severity biomarkers (serum LDH and β2 microglobulin). The degree of CD200 expression was significantly positively correlated to Treg percentage (r = 0.565, p < 0.01). Analysis of the CD200 negative patients had a better progression free survival (p = 0.032) and overall survival (p = 0.04) as compared with those positive for CD200 expression. These findings illustrate a clear correlation between myeloma cell CD200 expression level and the frequency of immunosuppressive Treg cells. In conclusion, increased expression of CD200, expansion of suppressive Treg cells and elevation of cytokines might have a role in MM progression in this cohort of patients. Copyright © 2015 John Wiley & Sons, Ltd.     

外泌体在多发性骨髓瘤治疗中潜在作用

目的作为细胞间信息的重要传播者,外泌体在肿瘤诊断、治疗和预后等方面起着重要作用。研究证实其相关抑制剂或者类似物等具有抗多发性骨髓瘤(multiple myeloma,MM)的作用。本研究旨在总结国内外研究中外泌体在MM治疗中的潜在作用。方法以"外泌体、多发性骨髓瘤、治疗"为关键词,检索2004-08-2019-01PubMed和万方数据库检索系统中的文献。纳入标准:(1)外泌体的生物学功能;(2)外泌体在MM治疗中作用的研究。经筛选符合要求的文献有49篇。结果MM来源的外泌体某些成分如miRNA、蛋白质、lncRNA等与正常人之间表达存在差异,可利用其抑制剂或类似物发挥抗MM的作用,从而提供潜在治疗靶点。肿瘤来源外泌体及其加工修饰产物可作为一种新的癌症疫苗发挥抗肿瘤免疫;且外泌体的天然纳米结构使其成为药物递送的载体,可用于疾病的治疗。但目前疫苗相关研究较局限,药物载体在肿瘤治疗的应用方面研究亦甚少。结论外泌体可通过特异性表达某些成分而提供潜在治疗靶点,如其本身及其加工产物可制成疫苗,或以运载药物的形式在MM的治疗中发挥潜在作用。但目前这些研究尚处于初级阶段,外泌体应用于MM临床治疗尚需大量研究进一步证实。     

靶向干预多发性骨髓瘤骨髓间充质干细胞的实验研究

目的:间充质干细胞(MSCs)对维持多发性骨髓瘤(MM)骨髓微环境的稳定有重要作用,靶向干预MSCs有望成为MM治疗新的途径,本实验通过分析沙利度胺(Thalidomide,Thal)和地塞米松(dexamethasone,Dex)对MM患者MSCs凋亡及其IL-6、IL-1β、SCF表达的影响,探讨上述药物可能新的作用机制和MSCs作为治疗靶点的可行性。方法:含10?S的1640培养液培养MSCs,以终浓度分别为0μmol/L、1.0μmol/L、3.0μmol/L、5.0μmol/L、7.0μmol/L的Thal,0μmol/L、0.1μmol/L、1.0μmol/L、5.0μmol/L、10.0μmol/L的Dex作用MSCs8h,倒置光学显微镜和AnnexinV/FITC-PI双染法荧光显微镜下观察细胞凋亡,RT-PCR检测分析IL-6、IL-1β和SCF细胞因子表达的变化。结果:Thal、Dex均以浓度依赖方式诱导MM患者MSCs凋亡;Thal下调MSCs三种细胞因子的表达(P<0.05),对IL-6的抑制随药物浓度增加而加重,对IL-1β和SCF表达的下调与药物浓度无明显关系(P>0.05);Dex明显抑制MSCsIL-6、IL-1β、SCF的表达(P<0.05),对3种细胞因子的抑制呈药物浓度依赖性。结论:Thal、Dex能诱导MM患者MSCs的凋亡,抑制其IL-6、IL-1β、SCF的表达,是有效治疗MM的靶点和机制之一。     

Prognostic factors associated with progression of smoldering multiple myeloma to symptomatic form

BACKGROUND:

Smoldering multiple myeloma (SMM) presents a high risk of progression to symptomatic MM (sy‐MM). Herein, we analyzed some predictors of development of sy‐MM. In 144 patients with SMM, we also compared the risk of progression predicted by bone marrow plasma cell (BMPC) involvement on the bone marrow biopsy (BMB) versus bone marrow aspirates (BMA).

METHODS:

From January 1980 to July 2010, 397 patients with SMM observed in 12 centers of the Multiple Myeloma GIMEMA (Gruppo Italiano Malattie EMatologiche dell'Adulto) Latium Working Group have been analyzed. At progression to sy‐MM, the severity of clinical presentation was graded according to the need of intensive supportive care.

RESULTS:

After a median follow‐up of 135 months, the cumulative incidence of progression rates to sy‐MM were 45%, 55%, and 75% at 10, 15, and 20 years, respectively. Hemoglobin ≤12.5 g/dL, monoclonal component ≥2.5 g/dL, and BMPC ≥60% were the only parameters negatively affecting the cumulative incidence of progression. In particular, 10 of 397 (2.5%) patients with BMPC ≥60% had a 5.6‐fold increased risk of fast progression (within 2 years), which occurred with severe clinical manifestations in 62% of cases. BMB was more sensitive for the detection of BMPC involvement, even though BMA was a more reliable indicator of a rapid progression to sy‐MM.

CONCLUSIONS:

The highest risk of rapid evolution to sy‐MM and the severity of clinical manifestation at the progression suggest that SMM patients with a BMPC ≥60% should be treated soon after diagnosis. Moreover, BMPC is a more reliable index for progression to sy‐MM if assessed by BMA. Cancer 2012. © 2012 American Cancer Society.     

microRNA 在多发性骨髓瘤中作用的研究进展简

微小RNA（microRNA,miRNA）是一类长度为19—25个核苷酸的非编码小RNA,对于生物体的生长和发育起到关键的调节作用。不同microRNA在多发性骨髓瘤（MM）中有不同程度的表达。分析miRNA在MM中的异常表达有助于阐明MM的发生、发展机制,为MM的诊断、治疗提供新的理论依据。     

The effects of bortezomib on bone disease in patients with multiple myeloma

Bortezomib has demonstrated substantial activity in the treatment of patients with multiple myeloma and is widely incorporated into treatment strategies across the different settings. It is interesting to note that data are accumulating to suggest that the activity of bortezomib extends beyond the tumor cell and microenvironment to encompass effects on bone metabolism. Indeed, data from both the preclinical and clinical settings have suggested that bortezomib directly stimulates osteoblast growth and differentiation, while also inhibiting osteoclast development and activity. Notably, in the clinical setting, the bone anabolic effects of bortezomib could be demonstrated by the healing of lytic lesions as noted in some patients. These results are of importance because bone disease is a hallmark of myeloma and therefore any agent that combines antimyeloma activity with positive effects on bone is of substantial interest. However, further studies are needed to establish how the agent should be used for the treatment of patients with bone disease. Cancer 2014;120:618–623 . © 2013 American Cancer Society.     

多发性骨髓瘤伴非骨旁髓外病变临床分析

目的:探讨多发性骨髓瘤伴非骨旁髓外病变的临床特点。方法:回顾性分析2013 年3月至2016 年3 月在西安交通大学第一附属医院血液科住院的多发性骨髓瘤伴非骨旁髓外病变患者26例（A组）、病例资料及随访较完整的无髓外侵犯患者130例（B组）及伴骨旁髓外病变患者10例（C组）的临床资料,比较三者临床特点及预后。结果:多发性骨髓瘤伴非骨旁髓外病变患者发生率为4.68%（26/556）,发生部位为睾丸、椎管硬膜外、皮肤等。伴骨旁髓外病变患者发病率为1.80％（10/556）,发生部位为肋骨附近胸壁和脊柱旁等。A组中6例为反复治疗不缓解,出现髓外侵犯,自确诊到髓外病变出现中位时间为 3（2~12）个月;9例为完全缓解后在骨髓复发或未复发状态出现髓外侵犯,自确诊到髓外病变出现中位时间为16.5（8~45）个月;11例为初诊时伴有髓外侵犯。A组中位生存时间为17.2个月,B组中位生存时间为37.3个月,C组中位生存时间为35.3个月。结论:多发性骨髓瘤伴非骨旁髓外侵犯在骨髓不缓解和缓解状态均可出现,不依赖骨髓微环境生长,是骨髓瘤预后不良因素。骨髓瘤伴骨旁髓外病变患者预后优于伴非骨旁髓外病变患者。     

Choosing treatment options for patients with relapsed/refractory multiple myeloma

Multiple myeloma (MM) is a clonal plasma cell disorder that is still incurable using conventional treatments. Over the last decade, advances in front-line therapy have led to an increase in survival, but there are still some doubts in the case of relapsed/refractory disease. We searched the PubMed database for articles on treatment options for patients with relapsed/refractory MM published between 1996 and 2013. These treatments included hematopoietic cell transplantation (HCT), rechallenges using previous chemotherapy regimens, and trials of new regimens. The introduction of new agents such as the immunomodulatory drugs (IMIDs) thalidomide and lenalidomide, and the first-in-its-class proteasome inhibitor bortezomib, has greatly improved clinical outcomes in patients with relapsed/refractory MM, but not all patients respond and those that do may eventually relapse or become refractory to treatment. The challenge is therefore to select the optimal treatment for each patient by balancing efficacy and toxicity. To do this, it is necessary to consider disease-related factors, such as the quality and duration of responses to previous therapies, and the aggressiveness of the relapse, and patient-related factors such as age, comorbidities, performance status, pre-existing toxicities and cytogenetic patterns. The message from the trials reviewed in this article is that the new agents may be used to re-treat relapsed/refractory disease, and that the sequencing of their administration should be modulated on the basis of the various disease and patient-related factors. Moreover, our understanding of the pharmacology and molecular action of the new drugs will contribute to the possibility of developing tailored treatment.     

Thalidomide in the treatment of multiple myeloma

Thalidomide – banned from clinical use in the 1960s because of severe teratogenicity – is now back in clinical practice as an effective agent in the treatment of relapsed and refractory multiple myeloma. Several clinical trials have determined that thalidomide is active in 25–35% of patients with relapsed myeloma. The role of thalidomide in early-stage myeloma is being actively investigated. Thalidomide has antiangiogenic and immunomodulatory properties and is an effective inhibitor of TNF-α. However, the mechanism of its action in myeloma remains unclear. Major toxicities of thalidomide include constipation, sedation, skin rash, fatigue and peripheral neuropathy. This paper summarizes the current status of thalidomide in multiple myeloma.     

多发性骨髓瘤完全缓解后微小残留病变的检测进展及意义

 已经证实传统意义上完全缓解（CR）的患者体内仍残留约1010 个肿瘤细胞,是疾病进展和复发的根源。为进一步了解体内残存的肿瘤细胞,聚合酶链反应（polymerase chain reaction,PCR）检测IgH基因单克隆重排、流式细胞术（flow cytometic,FCM）检查细胞表面抗原及染色体异常倍体和荧光原位杂交（flurescence in situ hybridizztion,FISH）分析染色体异常核型可用于临床以监测微小残留病的动态变化,判断预后并指导治疗。