Analysis of pigmented villonodular synovitis with genome‐wide complementary DNA microarray and tissue array technology reveals insight into potential novel therapeutic approaches

Objective

To characterize the gene expression profile and determine potential diagnostic markers and therapeutic targets in pigmented villonodular synovitis (PVNS).

Methods

Gene expression patterns in 11 patients with PVNS, 18 patients with rheumatoid arthritis (RA), and 19 patients with osteoarthritis (OA) were investigated using genome‐wide complementary DNA microarrays. Validation of differentially expressed genes was performed by real‐time quantitative polymerase chain reaction and immunohistochemical analysis on tissue arrays (80 patients with PVNS, 51 patients with RA, and 20 patients with OA).

Results

The gene expression profile in PVNS was clearly distinct from those in RA and OA. One hundred forty‐one up‐regulated genes and 47 down‐regulated genes were found in PVNS compared with RA, and 153 up‐regulated genes and 89 down‐regulated genes were found in PVNS compared with OA (fold change ≥1.5; Q ≤ 0.001). Genes differentially expressed in PVNS were involved in apoptosis regulation, matrix degradation, and inflammation (ALOX5AP, ATP6V1B2, CD53, CHI3L1, CTSL, CXCR4, HSPA8, HSPCA, LAPTM5, MMP9, MOAP1, and SPP1).

Conclusion

The gene expression signature in PVNS is similar to that of activated macrophages and is consistent with the local destructive course of the disease. The gene and protein expression patterns suggest that the ongoing proliferation in PVNS is sustained by apoptosis resistance. This result suggests the possibility of a potential novel therapeutic intervention against PVNS.

     

Successful arthroscopic treatment of pigmented villonodular synovitis of the knee in a patient with congenital deficiency of plasminogen activator inhibitor-1 and recurrent haemarthrosis.

We report the arthroscopic treatment of pigmented villonodular synovitis (PVNS) in a 13-year-old Japanese boy with congenital partial deficiency of plasminogen activator inhibitor-1 (PAI-1). He was admitted to our hospital with recurrent haemarthrosis of his right knee. Characteristic abnormalities of fibrinolysis included shortened euglobulin lysis time, low PAI-1 activity and low PAI-1 antigen levels. In addition, levels of "active PAI" in the plasma, which is a measure of total PAI bound to exogenous plasminogen activator, were very low. These parameters remained low after venous occlusion. The diagnosis of PVNS was established by synovial membrane biopsy, and arthroscopic synovectomy was performed with adjuvant administration of intravenous tranexamic acid. Subsequent bleeding episodes have been well controlled by oral administration of tranexamic acid on demand.     

Animal models of systemic sclerosis: insights into systemic sclerosis pathogenesis and potential therapeutic approaches

PURPOSE OF REVIEW: Animal models have been extremely valuable in contributing to a better understanding of the pathogenesis of systemic sclerosis. Discussed here are recent studies that have examined the molecular pathways and potential therapeutic approaches for systemic sclerosis using animal models. RECENT FINDINGS: Reported evidence further indicates that the immune system plays a role in modulating the fibrosis observed in the tight skin-1/+ mouse model for systemic sclerosis. CD19, interleukin-6, and interleukin-4 are involved. The injection of spleen cells into immune-compromised mice resulted in fibrotic, vascular, and immunologic alterations quite similar to those of systemic sclerosis. Transforming growth factor-beta and its signaling pathway (JAK kinase and STAT-6, Smad2/3, and Smad7) appear to play a central role in the development of fibrosis as well as monocyte chemoattractant protein-1, CCR-2, platelet-derived growth factor C, and excessive apoptosis. Viruses were shown to be possible cofactors. The therapeutic agents hepatocyte growth factor and halofuginone were shown to prevent fibrosis in animal models of systemic sclerosis. SUMMARY: The transforming growth factor-beta signaling pathway is a common mechanism of tissue fibrosis in animal models of systemic sclerosis, although numerous additional molecules modulate this pathway or have a direct effect on fibrosis.     

Treatment of pigmented villonodular synovitis with yttrium-90: Changes in immunologic features,Tc-99m uptake measurements,and MR imaging of one case

Summary Pigmented villonodular synovitis (PVNS) is an uncommon proliferative disease of synovium. We report a 35-year-old male with diffuse form of PVNS of left knee, treated with intraarticular injection of 5 mCi of yttrium-90 (Y-90) silicate colloid consisting of two doses with a 3-month interval between them. During follow-up, the affected knee showed clinical improvement and was accompanied by a decrease of the levels of soluble interleukin-2 receptor in sera and synovial fluids (SF). When compared to osteoarthritis subjects, SF lymphocyte subsets of this case before Y-90 therapy showed a lower CD4: CD8 cell ratio and absence of suppressor inducer cells (CD4+2H4+). The Tc-99m pertechnetate knee uptake indexes correlated well with clinical improvement. Serial magnetic resonance imaging revealed significant change one year after Y-90 therapy. The findings of immunological assessment suggested that immunoregulatory dysfunction may be related to the pathogenesis of PVNS.     

Coexpression network analysis coupled with connectivity map database mining reveals novel genetic biomarkers and potential therapeutic drugs for polymyositis

Clinical Rheumatology - Polymyositis (PM) is a chronic autoimmune connective tissue disease whose pathogenic mechanisms are unclear. This study aimed to identify the main genes and functionally...     

Treatment of pigmented villonodular synovitis of the knee with yttrium-90 silicate: prospective evaluations by arthroscopy, histology, and 99mTc pertechnetate uptake measurements.

The diffuse form of pigmented villonodular synovitis of eight knee joints of eight patients was treated by intra-articular injection of 185 MBq yttrium-90 silicate (90Y). Six patients had a recurrence of disease after one or two surgical synovectomies. After treatment with 90Y once or twice four knees showed clinical improvement with an accompanying decrease of the inflammatory activity as measured by the technetium-99m pertechnetate (99mTcO4-) uptake ratio and the severity of the diseased synovial tissue. Arthroscopy was performed before and six months after each 90Y treatment. The ratio of 99mTcO4- uptake in the inflamed compared with the normal knee joint correlated well with the macroscopical grading of pigmented villonodular synovitis. In all cases areas of persistent synovitis were found after the 90Y injection and this was confirmed both by histological examination and 99mTcO4- uptake measurements. Biopsy specimens taken from the diseased synovial areas showed histologically mostly less prominent and less numerous villi. The cartilage damage was slightly increased in only two cases. No radiological deterioration was found during follow up (mean 24 months, range 12-41). No complications of the radiosynoviortheses were noted.     

Data-independent acquisition-based quantitative proteomic analysis of m.3243A>G MELAS reveals novel potential pathogenesis and therapeutic targets

The pathogenesis of mitochondrial myopathy, encephalopathy, lactic acidosis and stroke like episodes (MELAS) syndrome has not been fully elucidated. The m.3243A > G mutation which is responsible for 80% MELAS patients affects proteins with undetermined functions. Therefore, we performed quantitative proteomic analysis on skeletal muscle specimens from MELAS patients. We recruited 10 patients with definitive MELAS and 10 age- and gender- matched controls. Proteomic analysis based on nanospray liquid chromatography-mass spectrometry (LC-MS) was performed using data-independent acquisition (DIA) method and differentially expressed proteins were revealed by bioinformatics analysis. We identified 128 differential proteins between MELAS and controls, including 68 down-regulated proteins and 60 up-regulated proteins. The differential proteins involved in oxidative stress were identified, including heat shock protein beta-1 (HSPB1), alpha-crystallin B chain (CRYAB), heme oxygenase 1 (HMOX1), glucose-6-phosphate dehydrogenase (G6PD) and selenoprotein P. Gene ontology and kyoto encyclopedia of genes and genomes pathway analysis showed significant enrichment in phagosome, ribosome and peroxisome proliferator-activated receptors (PPAR) signaling pathway. The imbalance between oxidative stress and antioxidant defense, the activation of autophagosomes, and the abnormal metabolism of mitochondrial ribosome proteins (MRPs) might play an important role in m.3243A > G MELAS. The combination of proteomic and bioinformatics analysis could contribute potential molecular networks to the pathogenesis of MELAS in a comprehensive manner.     

Revisiting the immune microenvironment of diffuse large B-cell lymphoma using a tissue microarray and immunohistochemistry: robust semi-automated analysis reveals CD3 and FoxP3 as potential predictors of response to R-CHOP

Gene expression studies have identified the microenvironment as a prognostic player in diffuse large B-cell lymphoma. However, there is a lack of simple immune biomarkers that can be applied in the clinical setting and could be helpful in stratifying patients. Immunohistochemistry has been used for this purpose but the results are inconsistent. We decided to reinvestigate the immune microenvironment and its impact using immunohistochemistry, with two systems of image analysis, in a large set of patients with diffuse large B-cell lymphoma. Diagnostic tissue from 309 patients was arrayed onto tissue microarrays. Results from 161 chemoimmunotherapy-treated patients were used for outcome prediction. Positive cells, percentage stained area and numbers of pixels/area were quantified and results were compared with the purpose of inferring consistency between the two semi-automated systems. Measurement cutpoints were assessed using a recursive partitioning algorithm classifying results according to survival. Kaplan-Meier estimators and Fisher exact tests were evaluated to check for significant differences between measurement classes, and for dependence between pairs of measurements, respectively. Results were validated by multivariate analysis incorporating the International Prognostic Index. The concordance between the two systems of image analysis was surprisingly high, supporting their applicability for immunohistochemistry studies. Patients with a high density of CD3 and FoxP3 by both methods had a better outcome. Automated analysis should be the preferred method for immunohistochemistry studies. Following the use of two methods of semi-automated analysis we suggest that CD3 and FoxP3 play a role in predicting response to chemoimmunotherapy in diffuse large B-cell lymphoma.     

Invariant natural killer T (iNKT) cells in asthma: a novel insight into the pathogenesis of asthma and the therapeutic implication of glycolipid ligands for allergic diseases.

Allergic bronchial asthma is a complex inflammatory diseases originated from dysregulated immune responses in the respiratory mucosa. The inflammatory state in asthmatic lung is characterized by massive infiltration with eosinophils, lymphocytes, and mast cells in the airway mucosa leading to airway hyperseisitivity, goblet cell hyperplasia and mucus overproduction. The inflammatory process is thought to be the result of intensive T helper (Th) 2-biased immune response. Over the past several years, there has been enormous progress in understanding the mechanisms for development of Th2-biased responses after inhaled exposure to allergens and the characteristics of CD4+ T cells prominently involved in this process. Recently, a new population of T cells, invariant natural killer T (iNKT) cells has been shown to play an important role in the pathogenesis of mouse model of allergic airway inflammation. iNKT cells are one of the most potent immune modulators through a massive production of a various cytokines including IL-4 and IFN-gamma upon activation, and are involved in a variety of immunoregulations including infection, autoimmunity, and tumor surveillance. The potent pathogenic role of iNKT cells in the development of bronchial asthma is due to their ability to produce predominant Th2 cytokines in a given condition. The involvement of iNKT cells in the pathogenesis of asthma might have been underestimated in the past studies demonstrating the involvement of CD4+ T cells in asthma because of the difficulty in the detection of iNKT cells. Meanwhile, growing evidences have demonstrated that iNKT cells could be a promising target for immune-based therapies for autoimmune diseases, tumor, and infection due to the invariance of their TCR usage, the restriction to the evolutionally-conserved non-polymorphic antigen-presenting molecule CD1d, and their outstanding ability to produce both Th1- and Th2-cytokines. In this review, we will overview current understanding of the pathophysiological roles of iNKT cells in asthma. We would also discuss on possible therapeutic approaches to bronchial asthma employing glycolipid ligands for iNKT cells.     

The interdependence of skin aging, skin cancer, and DNA repair capacity: a novel perspective with therapeutic implications

The human body is constantly exposed to exogenous and endogenous insults that threaten its genomic integrity and that lead to changes at the molecular, biochemical, and cellular levels. As a major interface between the environment and the internal milieu, our skin is especially subject to such events. Common insults include but are not limited to infectious agents, environmental pollutions and toxins, carcinogens, and ultraviolet (UV) irradiation. It is estimated that there are thousands of DNA alterations in each cell daily. Therefore, if not efficiently repaired, our genome would rapidly be destroyed. This review focuses predominantly on UV-induced DNA damage in human skin, protective molecular responses to UV damage, and the consequences of these opposing forces for aging and photocarcinogenesis.     

Analysis of circulatory mitochondrial DNA level after cardiac surgery with cardiopulmonary bypass and potential prognostic implications

Our research letter found that circulatory mtDNA level increased after the end of CPB and positive correlations between mtDNA and peak CRP level, peak BNP level, and peak PCT level, which revealed the prognostic role of perioperative circulatory mtDNA level in patients who underwent cardiopulmonary bypass.     

Excretion into bile of a novel unenveloped DNA virus (TT virus) associated with acute and chronic non-A-G hepatitis

Recently, an unenveloped, single-stranded DNA virus named TT virus (TTV) has been reported in association with hepatitis of non-A-G etiology. Five patients with TTV viremia, who received bile drainage or cholecystectomy, were tested for TTV DNA in bile by polymerase chain reaction with heminested primers. TTV DNA was detected in bile from all patients; titers were 10-100 times higher than in serum in 4 and at a comparable level in the remaining 1 patient. TTV DNA was detected in feces, also, in 1 of the 2 patients tested. The buoyant density of TTV in bile from 1 tested patient (1.33-1.35 g/cm3) was the same as that in feces (1.32-1.35 g/cm3). TTV may be secreted via bile into feces in a transmissible form and would spread by a fecal-oral route for deep and wide penetration into the general population.     

Analysis of the murine All-1 gene reveals conserved domains with human ALL-1 and identifies a motif shared with DNA methyltransferases.

A series of translocation break points found in a subset of human acute leukemias have one of the breaks on human chromosome 11q23. This region has recently been cloned and a large gene, ALL-1, with homology to the Drosophila trithorax gene has been identified. This paper describes the cloning, sequencing, and mapping of the mouse homolog of ALL-1. We have found a motif present in All-1 that shows homology to the zinc-binding domain of DNA (cytosine-5) methyltransferases (EC 2.1.1.63). Sequence analysis of the murine All-1 gene has identified distinct regions of homology with the human ALL-1 gene; these highly conserved domains may define regions of functional significance in mammals. In addition, we have identified alternatively spliced forms of All-1 within one of the zinc-finger domains, suggesting that there may be different targets and/or functions for All-1 proteins. Finally, we report that All-1 resides in the proximal portion of mouse chromosome 9 and is a candidate for a mutation that results in skeletal transformations during embryonic development.     

Mice with targeted disruption of <Emphasis Type="Italic">p8</Emphasis>gene show increased sensitivity to lipopolysaccharide and DNA microarray analysis of livers reveals an aberrant gene expression response

Background  

p8 is a DNA-binding protein induced in many tissues in response to LPS treatment. Hence, p8 could be a mediator of LPS-associated effects or, on the contrary, p8 expression may be part of the protective mechanism of the tissues in response to LPS. Finally, p8 expression in response to LPS could also be a simple epiphenomenon.     

血吸虫生殖产卵相关基因pcDNA3／Sj SDISPDNA在小鼠体内的组织分布及整合的观察

目的探讨日本血吸虫生殖产卵相关基因DNA在小鼠体内组织分布以及整合到宿主基因组上的可能性。方法将生殖产卵相关基因重组喷柱pcDNA3／SjSDISP经肌注免疫小鼠后,在注射后12b、1w、3w和6w取小鼠各组织,抽提其基因组DNA进行PCR扩增,然后纯化基因组DNA进行PCR扩增,结果经琼脂糖凝胶电泳分析。结果注射后12h,在血液、心、肝、脾、肺、肾及注射鄂位肌肉中可检测到pcDNA3／Sj SDISP,至第1w,仍可在部分小鼠中检测到,至第3w不再检出。未发现生殖产卵相关基因pcDNA3／Sj SDISP整合入小鼠基因组中。结论肌注pcDNA3／Sj SDISP后,可在组织中广泛分布,但并未持续存在体内。未发现与宿主细胞基因组整合的直接证据