Targeting tumor micro-environment for design and development of novel anti-angiogenic agents arresting tumor growth

Angiogenesis: a process of generation of new blood vessels has been proved to be necessary for sustained tumor growth and cancer progression. Inhibiting angiogenesis pathway has long been remained a significant hope for the development of novel, effective and target orientated antitumor agents arresting the tumor proliferation and metastasis. The process of neoangiogenesis as a biological process is regulated by several pro- and anti-angiogenic factors, especially vascular endothelial growth factor, fibroblast growth factor, epidermal growth factor, hypoxia inducible factor 1 and transforming growth factor. Every endothelial cell destined for vessel formation is equipped with receptors for these angiogenic peptides. Moreover, numerous other angiogenic cytokines such as platelet derived growth factor (PGDF), placenta growth factor (PGF), nerve growth factor (NGF), stem-cell factor (SCF), and interleukins-2, 4, 6 etc. These molecular players performs critical role in regulating the angiogenic switch. Couple of decade's research in molecular aspects of tumor biology has unraveled numerous structural and functional mysteries of these angiogenic peptides. In present article, a detailed update on the functional and structural peculiarities of the various angiogenic peptides is described focusing on structural opportunities made available that has potential to be used to modulate function of these angiogenic peptides in developing therapeutic agents targeting neoplastic angiogenesis. The data may be useful in the mainstream of developing novel anticancer agents targeting tumor angiogenesis. We also discuss major therapeutic agents that are currently used in angiogenesis associated therapies as well as those are subject of active research or are in clinical trials.     

Multiple myeloma in a murine syngeneic model:modulation of growth and angiogenesis by a monoclonal antibody to kininogen

Multiple myeloma (MM), a B-cell malignancy characterized by proliferation of monoclonal plasma cells remains incurable. Murine plasma cell tumors share common features with human MM. We used two cell lines (B38 and C11C1) derived from P3X63Ag8 myeloma cells. The new cell lines were implanted subcutaneously in the strain of origin (Balb/c mice) and used as a model to monitor the effects of C11C1 monoclonal antibody (mAb) to kininogen (HK). We assessed their behavior by intraperitoneal and subcutaneous implantation, by implanting them together and by treating B38–MM with purified mAb C11C1. We evaluated growth, microvascular density (MVD), and cellular expression of urokinase-type plasminogen activator-receptor (uPAR), fibroblast growth factor-2 (FGF-2), vascular endothelial growth factor (VEGF), bradykinin-1 receptor (B1R), bradykinin-2 receptor (B2R) and HK. We found that both MM-cell-lines are uPAR positive, that mAb C11C1 inhibits its own tumor growth in vivo, slows down B38-MM growth rate when both MM are implanted together and when mAb C11C1 is injected intraperitoneally. MAb C11C1-treated-MM showed decreased MVD and HK binding in vivo without FGF-2, B1R or B2R expression changes. We propose that the B38-extramedullary-myeloma-model is a useful tool to study the interactions of this hematopoietic tumor and its environment and that mAb C11C1 may improve the efficacy of conventional MM treatment with minimal side effects. Financial support: NIH grants. R01 CA-083121-08 (R.W.C.) and T32 HL-07777-14 (R.W.C.)     

Frequent detection of Human Herpes Virus-8 in bone marrow of Jordanian patients of multiple myeloma

The association between Human Herpes Virus-8 (HHV-8), also called Kaposi's sarcoma associated herpesvirus (KSHV), and the pathogenesis of multiple myeloma remains controversial. Many past studies conducting on different populations have come to contradicting conclusions. In this study, we attempted to investigate the presence of HHV-8 in Jordanian multiple myeloma patients. We carried out nucleic acid amplification reactions targeting specific viral DNA sequences on 35 fresh bone marrow aspirate samples from 17 patients with multiple myeloma, 9 patients with various hematological malignancies and 9 normal subjects. HHV-8 specific sequences were detected in 7 out of 17 multiple myeloma patients (41%) using primers specific for the open reading frame region 26 (ORF26). All patients with other hematological malignancies as well as the normal subjects did not harbour the virus. These findings support the previous reports of frequent detection of HHV-8 in bone marrow of multiple myeloma patients.     

Sex hormones and immune dysregulation in multiple myeloma

A group of 49 multiple myeloma patients, 20 men and 29 women, were evaluated. Follicle-stimulating hormone (FSH), luteinizing hormone (LH), 17-oestradiol (E) and testosterone (T) serum concentrations have been detected by radioimmunoassay. Peripheral blood lymphocyte proliferation in response to phytohaemagglutinin (PHA), concanavalin A (ConA), recombinant interleukin-2 (rIL-2) and dextran sulphate (DxS) was investigated. Our findings provide evidence for two different patterns of sex hormone changes and immune dysfunctions presented differently by male and female multiple myeloma patients. In men increased FSH, LH and E concentrations and an augmented E to T ratio were associated with decreased lymphocyte blastogenic response to PHA, ConA and increased proliferation to rIL-2 and DxS. Female patients with multiple myeloma demonstrated normal values of FSH, LH and T, but a diminished E level and decreased E to T ratio correlated with a lymphocyte normal response to PHA and ConA and augmented blastogenesis to IL-2 and DxS. Our data, while admittedly preliminary, suffice to provide an indication of sex hormone changes in multiple myeloma patients, which could be responsible, at least in part, for the immune dysfunction observed in multiple myeloma.     

Modification of the bone marrow MSC population in a xenograft model of early multiple myeloma

Multiple myeloma (MM) is a hematological malignancy characterized by clonal proliferation of abnormal plasma cells. MM dysregulates the homeostasis of the bone niche cells like osteoclasts and osteoblasts, responsible for the bone maintenance leading to bone loss and hypercalcemia, as well as the normal immune cells leading to immunodeficiency and anemia. Osteoblasts are part of the cell population differentiating from mesenchymal stem cells (MSC). MSC also gives rise to other cell types such as adipocytes and chondrocytes. It has been observed that adipocytes support MM growth by increasing its survival and chemo-resistance. As adipocytes originate from MSC, the understanding of early modifications in the MSC population during the disease progression is of paramount importance and may help for early diagnosis of MM. Herein, we have evaluated the modification of the MSC population in the bone niche in an in vivo model of MM. Our results showed that before an observable engraftment of MM in the bone niche, the proportion of MSC population is significantly decreased, while a significant increase in adipocyte related genes such as PPARγ and CEBPα expression appears, with no difference in osteogenic differentiation. These results suggest that the bone niche is switching to a “fatty” marrow which would create an adequate microenvironment for MM. This led us to screen for and identify modulated adipokines in the sera of this in vivo MM-mice model. Such changes could reflect early signs of MM and potentially be exploited as detection biomarkers of the disease.     

A novel panel of protein biomarkers for predicting response to thalidomide-based therapy in newly diagnosed multiple myeloma patients

Multiple myeloma (MM) is a heterogeneous group of disorders both genotypically and phenotypically. Response to thalidomide-based induction therapy in newly diagnosed patients varies significantly in published clinical trials. Proteomic analysis was performed on 39 newly diagnosed MM patients treated with a thalidomide-based regimen (22 responders; 17 non-responders) using immunodepletion, 2-D DIGE analysis and mass spectrometry. Zinc-α-2-glycoprotein (ZAG), vitamin D-binding protein (VDB), serum amyloid-A protein (SAA) and β-2-microglobulin (B2M) had statistically significant higher concentrations in non-responders compared to responders, while haptoglobin (Hp) had a lower concentration. ELISAs were used to validate the candidate protein biomarkers using unfractionated serum from 51 newly diagnosed MM patients (29 responders; 22 non-responders). Using logistic regression, the best possible area under the curve (AUC) was 0.96 using ZAG, VDB and SAA in combination. Leave-one-out-cross-validation (LOOCV) indicated an overall predictive accuracy of 84% with associated sensitivity and specificity values of 81.8 and 86.2%, respectively. Subsequently, 16 of 22 thalidomide-refractory patients successfully achieved complete response or very good partial response using second-line treatment suggesting that the biomarker profile is specific to thalidomide response rather than identifying patients with MM refractory to all therapies. Using a novel panel of predictive biomarkers, the feasibility of predicting response to thalidomide-based therapy in previously untreated MM has been demonstrated.     

多发性骨髓瘤患者医院感染临床分析

目的 探讨多发性骨髓瘤患者医院感染的临床状况和危险因素。方法 对1994年1月～2001年12月收治的62例多发性骨髓瘤患者医院感染发生情况进行调查分析。结果 多发性骨髓瘤患者发生医院感染33例。感染发生率为53．2％,好发部位为呼吸道、口腔、胃肠道和泌尿道等。结论 多发性骨髓瘤患者医院感染的发生与骨髓瘤细胞含量、疾病Ⅲ期、骨损害、白细胞减少、化疗及住院日等因素有关。应采取相应措施,避免医院感染发生。     

The role of microenvironment in tumor angiogenesis

Tumor microenvironment is essential for tumor cell proliferation, angiogenesis, invasion and metastasis through its provision of survival signals, secretion of growth and pro-angiogenic factors, and direct adhesion molecule interactions. This review examines its importance in the induction of an angiogenic response in tumors and in multiple myeloma. The encouraging results of pre-clinical and clinical trials in which tumors have been treated by targeting the tumor microenvironment are also discussed.     

免疫固定电泳在诊断多发性骨髓瘤中的作用

目的 探讨分析免疫固定电泳在诊断多发性骨髓瘤（MML）中的作用。方法 以2014年10月至2016年10月丽水市人民医院收治的110例需进行免疫固定电泳检测的患者为研究对象,对其进行血清M蛋白和浓缩尿中M蛋白的检测,筛选出两种检测均显阳性的患者,并将检测结果与临床资料相比较,对比血清M蛋白和浓缩尿中M蛋白的检测阳性与多发性骨髓瘤之间的关系。结果 110例需进行免疫固定电泳检测的患者中,经临床资料分析,有34例患者为多发性骨髓瘤。这34例患者血清M蛋白的检测显阳性,临床诊断符合率为100.00%;25例患者浓缩尿中M蛋白的检测显阳性,临床诊断符合率为73.53%。血清检验的临床符合率较尿液检验更好,差异具有统计学意义（P＜0.05）;34例血清M蛋白的检测显阳性的患者中,IgG型21例（61.76%）,IgA型8例（23.53%）,IgM型3例（8.82%）,γ轻链型2例（5.88%）。结论 MML患者采用血清及尿液免疫固定电泳检测,血清M蛋白检测阳性率较浓缩尿中M蛋白的检测阳性率要高,血清及尿液进行的免疫固定电泳检测对MML的诊断、检测及预后判断具有重要的提示作用,临床工作者可以根据电泳图谱对MML进行分型,并制定个体化的治疗方案。     

Degrasyn-like symmetrical compounds: Possible therapeutic agents for multiple myeloma (MM-I)

A series of degrasyn-like symmetrical compounds have been designed, synthesized, and screened against B cell malignancy (multiple myeloma, mantle cell lymphoma) cell lines. The lead compounds T5165804 and CP2005 showed higher nanomolar potency against these tumor cells in comparison to degrasyn and inhibited Usp9x activity in vitro and in intact cells. These observations suggest that this new class of compounds holds promise as cancer therapeutic agents.     

Glycosylation of polyclonal and paraprotein IgG in multiple myeloma

It has previously been shown that in multiple myeloma (MM) each IgG paraprotein exhibits a unique oligosaccharide profile. It has been assumed that this results from a clone specific glycosylation machinery. However, the abnormal physiological environment of the bone marrow in this disease may also affect normal plasma cells producing polyclonal IgG. We present data to show that this is so and that, in two cases, the oligosaccharide profile of the polyclonal IgG reflected that of the paraprotein from the same patient rather than that of normal polyclonal IgG. This revised version was published online in November 2006 with corrections to the Cover Date.     

多发性骨髓瘤预后相关自噬基因筛选及建模

为探讨自噬相关基因(ARGs)在MM发生发展中的作用机制并建立相关的预后模型。基于MMRF与HADb数据库,通过R语言确定多发性骨髓瘤中自噬相关基因的差异表达,GO和KEGG分析自噬相关基因与多发性骨髓瘤发生发展的关系,使用COX回归算法建立多基因预后模型,Kaplan-Meier方法绘制生存曲线,ROC曲线评价预后模型的可靠性。最终从764例多发性骨髓瘤患者骨髓样本及4例正常骨髓样本中共发现104个基因的表达在多发性骨髓瘤样本中具有显著差异,其中上调基因46个,下调基因58个。GO富集主要集中在巨自噬、自噬调节、细胞对外部刺激的反应等本体学注释。KEGG富集主要集中在自噬、细胞凋亡、NOD样受体信号通路、PI3K-Akt信号通路。单因素COX分析发现33个自噬相关基因与多发性骨髓瘤患者整体生存明显相关。多因素COX回归筛选出13个预后相关自噬相关基因(NKX2-3、NCKAP1、BIRC5、PEX3、HGS、RUBCN、PARP1、ARSA、DNAJB9、HSP90AB1、EEF2、FKBP1B和CD46)建立多发性骨髓瘤自噬相关基因预后模型。Kaplan-Meier生存曲线分析显示...     

Geospatial analysis of population-based incidence of multiple myeloma in the United States

BackgroundWe investigated the spatial patterns of multiple myeloma (MM) incidence in the United States (US) between 2013 and 2017 to improve understanding of potential environmental risk factors for MM.MethodsWe analyzed the average county-level age-adjusted incidence rates (“ASR”) of MM between 2013 and 2017 in 50 states and the District of Columbia using the U.S. Cancer Statistics Public Use Databases. We firstly divided the ASR into quintiles and described spatial patterns using a choropleth map. To identify global and local clusters of the ASR, we performed the Spatial Autocorrelation (Global Moran’s I) analysis and the Anselin’s Local Indicator of Spatial Autocorrelation (LISA) analysis. We compared the means of selected demographic and socioeconomic factors between the clusters and counties of the whole US using Welch one-sided t-test.ResultsWe identified distinct spatial dichotomy of the ASR across counties. High ASR were observed in counties in the Southeast of the US as well as the Capital District (metropolitan areas surrounding Albany) and New York City in the state of New York, while low ASR were observed in counties in the Southwest and West of the US. The ASR showed a significant positive spatial autocorrelation. We identified two major high-high local clusters of the ASR in Georgia and Southern Carolina and five major low-low local clusters of the ASR in Alabama, Arizona, New Hampshire, Ohio, Oregon, and Tennessee. The racial population distribution may partly explain the spatial distribution of MM incidence in the US.ConclusionFindings from this study showed distinct spatial distribution of MM in the US and two high-high and five low-low local clusters. The non-random distribution of MM suggests that environmental exposures in certain regions may be important for the risk of MM.     

Incidence of multiple myeloma in Kailuan cohort: A prospective community-based study in China

BackgroundPrevious retrospective studies showed that the incidence and mortality rates for MM in China were lower than those in western countries. A large-scale prospective study on incidence and mortality rates of MM is still lacking.MethodsBased on the prospective Kailuan Cohort study in China, we included all patients with MM in Kailuan Cohort from June 1, 2008 to December 31, 2016. Using the numbers of diagnosed cases and deaths during the study period as the numerators and the corresponding observed person-years as the denominators respectively, we calculated crude incidence and mortality rates. The 95% confidence intervals for crude incidence rate and mortality rate were estimated base on Poisson distribution. Rates were standardized by direct standardization according to the China population in 2000 and Segi’ world standard population.ResultsA total of 22 members from Kailuan Cohort were first diagnosed with MM between 2008 and 2016. The calculated crude incidence rates were 2.8 (95% CI, 1.7–4.2) per 100,000 person-years for all participants. The standardized incidence rate was 0.9 per 100,000 person-years (95% CI, 0.5–2.1) when standardized by 2000 China population census data, and 1.0 per 100,000 person-years (95% CI, 0.6–1.8) when standardized by Segi’s world standard population (WSP). The calculated crude mortality rates were 2.3 (95% CI, 1.4–3.6) per 100,000 person-years. The mortality standardized by 2000 China population census data was 0.7 per 100,000 person-years (95% CI, 0.3–1.9), and 0.9 per 100,000 population (95% CI, 0.5–1.7) when standardized by Segi’s WSP. Both incidence and mortality for males were higher than that for females almost in all age groups. Both rates increased steadily with age.ConclusionIn this community-based prospective cohort study, we found that the incidence of MM in China was far lower than that in American and Europe.     

Expression and pathogenesis of VCAM-1 and VLA-4 cytokines in multiple myeloma

ObjectiveThe objective of this study is to investigate the expression of Vascular cell adhesion molecule-1 (VCAM-1) and very late appearing antigen-4 (VLA-4) cytokines in MM (multiple Myeloma).MethodForty patients with MM are selected as the experimental group and 30 healthy persons as the control group. Flow cytometry is used to detect the expression of VCAM-1 (CD106), VLA-4 (CD49d), CD38 and CD138 antigens in experimental group and control group. ELISA (Enzyme Linked Immunosorbent Assay) is used to detect the concentration of VCAM-1 in serum of experimental group and control group. RT-PCR is used to detect the expression of VCAM-1.ResultsThe positive rate and antigen expression rate of VACM-1 antigen in the experimental group were significantly higher than those in the control group (P < 0.05). There were statistical differences of VLA-4 and VCAM-1 antigens between the initial diagnosis group and the relapse/refractory group, and between the relapse/refractory group and the platform stage group (P < 0.05). There were significant differences between VLA-4 antigen and VACM-1 antigen, phase I and phase II, and between phase I and phase III (P < 0.05). The concentration of VCAM-1 and the expression of VCAM-1 mRNA in the experimental group were significantly higher than (P < 0.01). In the different stages of ISS (International Staging System) and different disease groups in the experimental group, the concentration of VCAM-1 and the expression level of VCAM-1 mRNA are significantly different among the three groups of stage I, II and III (P < 0.01). There is a significant difference between the initial diagnosis group, the relapse/refractory group and the platform group (P < 0.05).ConclusionThere are abnormal expressions of adhesion molecules VCAM-1 and VLA-4 in multiple myeloma patients, which are related to ISS staging.     

华蟾素胶囊对多发性骨髓瘤化疗患者的效果及对细胞免疫和肠道菌群的影响

目的 探讨华蟾素胶囊对多发性骨髓瘤(MM)化疗患者的临床疗效和对其细胞免疫、肠道细菌的影响。 方法 选取95例本院收治的MM化疗患者,通过随机分组分为观察组50例和对照组45例。对照组患者常规化疗方案,观察组在其基础上给予华蟾素胶囊治疗,两组治疗时间均持续3个疗程。对比治疗后MM化疗患者临床疗效、治疗前后细胞免疫指标水平差异和肠道细菌分布情况。 结果 两组疗效等级分布差异显著(P0.05),治疗后观察组CD4+/CD8+T、DC1/DC2、CD8+CD25+/CD3+T、Th1/Th2、CD4+CD25+/CD3+T水平相近(P>0.05),对照组各个细胞免疫指标均显著下降(均P0.05),治疗后,观察组这两个指数均升高(P结论 华蟾素胶囊联合化疗可以提高MM患者细胞免疫功能,改善肠道细菌平衡,具有较好的临床疗效。     

Assessment of serum thiol/disulfide homeostasis in multiple myeloma patients by a new method

Objectives: The etiology of multiple myeloma (MM) is not exactly known. This study investigated the role of thiol/disulfide homeostasis in the etiopathogenesis of MM.

Methods: Some 50 patients with MM (aged 39–84 years) and 50 sex-matched healthy volunteer controls (aged 50–91 years) participated in this study. Venous blood samples were collected, and levels of native thiols, total thiols, and disulfide were measured.

Results: Native and total thiol levels in the control group were determined to be higher than in the study and patient groups (P<0.001). Disulfide levels were found to be higher in the control group than in the study group and higher in newly diagnosed patients than in outpatients who were undergoing treatment (P=0.002). The ratios of thiol levels were found to be similar in both the study and control groups (P>0.05).

Discussion: The results of the study show that although there was a decrease in the levels of disulfide, native thiol, and total thiol, the balance of thiol/disulfide was maintained. This is the first study to research the homeostasis of dynamic thiol/disulfide from the perspective of the new method that was used. We hope that this study will encourage and facilitate further studies in this area.     

Communication between bone marrow mesenchymal stem cells and multiple myeloma cells: Impact on disease progression

Multiple myeloma (MM) is a hematological malignancy characterized by the accumulation of immunoglobulin-secreting clonal plasma cells at the bone marrow (BM). The interaction between MM cells and the BM microenvironment, and specifically BM mesenchymal stem cells (BM-MSCs), has a key role in the pathophysiology of this disease. Multiple data support the idea that BM-MSCs not only enhance the proliferation and survival of MM cells but are also involved in the resistance of MM cells to certain drugs, aiding the progression of this hematological tumor. The relation of MM cells with the resident BM-MSCs is a two-way interaction. MM modulate the behavior of BM-MSCs altering their expression profile, proliferation rate, osteogenic potential, and expression of senescence markers. In turn, modified BM-MSCs can produce a set of cytokines that would modulate the BM microenvironment to favor disease progression. The interaction between MM cells and BM-MSCs can be mediated by the secretion of a variety of soluble factors and extracellular vesicles carrying microRNAs, long non-coding RNAs or other molecules. However, the communication between these two types of cells could also involve a direct physical interaction through adhesion molecules or tunneling nanotubes. Thus, understanding the way this communication works and developing strategies to interfere in the process, would preclude the expansion of the MM cells and might offer alternative treatments for this incurable disease.