Jingjing Ding

Discoidin domain receptor 1 is associated with poor prognosis of non-small cell lung cancer and promotes cell invasion via epithelial-to-mesenchymal transition

Discoidin domain receptors (DDRs) are a novel class of receptor tyrosine kinases that respond to several collagens and facilitate cell adhesion. DDR1 is highly expressed in a variety of human cancers, and it is clear that DDR1 is primarily expressed in epithelial cells including lung, colon and brain. Moreover, DDR1 expression can be stimulated by collagen types I, II, III, IV, V, VIII and XI, and aberrant signaling induced by DDR1 dysregulated expression is involved in various steps of tumorigenesis. However, the molecular mechanism underlying the role of DDR1 in cancer development is not well documented. In this study, we found that the expression of DDR1 is upregulated in non-small cell lung cancer (NSCLC) tissues and cells when compared with counterpart normal tissues and cells. Furthermore, collagen I could induce DDR1 expression, and activated DDR1 promoted NSCLC cell migration and invasion, while knockdown of DDR1 by transfection with siRNA resulted in a significant decrease in cell migrativeness and invasiveness. Enhanced DDR1 expression mediated by collagen I could activate MMP-2, N-cadherin and vimentin expression, but reduce E-cadherin expression; however, inhibition of DDR1 expression could suppress MMP-2, N-cadherin and vimentin expression and induce E-cadherin activation. In conclusion, our findings indicated that upregulation of DDR1 induced by collagen I may contribute to the development and progression of NSCLC and this effect may be associated with increased invasiveness, at least in part, via promoting epithelial-to-mesenchymal transition.

Identification of novel driver mutations of the discoidin domain receptor 2 (DDR2) gene in squamous cell lung cancer of Chinese patients

BackgroundAlthough many of the recently approved genomically targeted therapies have improved outcomes for patients in non–small-cell lung cancer (NSCLC) with lung adenocarcinoma, little is known about the genomic alterations that drive lung squamous cell cancer (SCC) and development of effective targeted therapies in lung SCC is a promising area to be further investigated. Discoidin domain receptor 2 (DDR2), is a novel receptor tyrosine kinases that respond to several collagens and involved in tissue repair, primary and metastatic cancer progression.MethodsExpression of DDR2 mRNA was analyzed in 54 lung SCC tissues by qRT-PCR. Over-expression approaches were used to investigate the biological functions of DDR2 and its’ mutations in lung SCC cells. Conventional Sanger sequencing was used to investigate the mutations of DDR2 gene in 86 samples. The effect of DDR2 and its’ mutations on proliferation was evaluated by MTT and colony formation assays; cell migration and invasion was evaluated by trasnwell assays. Lung SCC cells stably transfected with pEGFP-DDR2 WT, pEGFP-DDR2-S131C or empty vector were injection into nude mice to study the effect of DDR2 and its’ mutation on tumorigenesis in vivo. Protein and mRNA expression levels of E-cadherin and MMP2 were determined by qRT-PCR and western blot analysis. Differences between groups were tested for significance using Student’s t-test (two-tailed).ResultsIn this study, we found that DDR2 mRNA levels were significantly decreased in 54 lung SCC tissues compared with normal lung tissues. Moreover, there were 3 novel DDR2 mutations (G531V, S131C, T681I) in 4 patients and provide the mutation rate of 4.6% in the 86 patients with lung SCC. The mutation of S131C in DDR2 could promote lung SCC cells proliferation, migration and invasion via inducing MMP-2, but reducing E-cadherin expression.ConclusionsThese data indicated that the novel DDR2 mutation may contribute to the development and progression of lung SCC and this effect may be associated with increased proliferation and invasiveness, at least in part, via regulating E-cadherin expression.

Meta-Analysis of the Adverse Effects of Long-Term Azithromycin Use in Patients with Chronic Lung Diseases

ABSTRACT The adverse effects of azithromycin on the treatment of patients with chronic lung diseases (CLD) were evaluated in the present study. MEDLINE and other databases were searched for relevant articles published until August 2013. Randomized controlled trials that enrolled patients with chronic lung diseases who received long-term azithromycin treatment were selected, and data on microbiological studies and azithromycin-related adverse events were abstracted from articles and analyzed. Six studies were included in the meta-analysis. The risk of bacterial resistance in patients receiving long-term azithromycin treatment was increased 2.7-fold (risk ratio [RR], 2.69 [95% confidence interval {95% CI}, 1.249, 5.211]) compared with the risk in patients receiving placebo treatment. On the other hand, the risk of bacterial colonization decreased in patients receiving azithromycin treatment (RR, 0.551 [95% CI, 0.460, 0.658]). Patients receiving long-term azithromycin therapy were at risk of increased impairment of hearing (RR, 1.168 [95% CI, 1.030, 1.325]). This analysis provides evidence supporting the idea that bacterial resistance can develop with long-term azithromycin treatment. Besides the increasingly recognized anti-inflammatory role of azithromycin used in treating chronic lung diseases, we should be aware of the potential for adverse events with its long-term use.

The overexpression of glypican-5 promotes cancer cell migration and is associated with shorter overall survival in non-small cell lung cancer

Although the correlation between glypican-5 (GPC5) and lung cancer is well known, the effect of GPC5 expression on non-small cell lung cancer (NSCLC) survival remains to be determined. In the present study, GPC5 expression in A549, H3255, and SPC-A1 NSCLC cell lines was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis. GPC5 mRNA and protein expression levels were found to be higher in A549 and H3255 cells compared with SPC-A1 cells. The role of GPC5 in NSCLC cell migration was evaluated in vitro by shRNA-mediated knockdown or the overexpression of GPC5 through scratch and transwell assays. The mean migration rates of cancer cells transfected with pRNAT-shRNA-GPC5-1 were reduced compared with the controls in A549 (P<0.001) and H3255 (P=0.001), while the migration rate of SPC-A1 with GPC5 overexpression was higher than that of the control (P=0.001). The downregulation of GPC5 impeded the transmigration of A549 and H3255 while the upregulation of GPC5 expression promoted the transmembrane invasion of SPC-A1. Furthermore, a panel of formalin-fixed paraffin-embedded NSCLC tissues from 127 patients undergoing curative resection (stages I, II and III) between January, 2003 and December, 2008 were obtained in order to investigate the correlation between GPC5 expression and clinicopathological factors using immunohistochemical methods. The results demonstrated that high GPC5 expression levels in NSCLC were associated with respiratory symptoms in lung cancer diagnosis, poor differentiation, vascular invasion, regional lymph node metastasis and a higher TNM stage. Using the Kaplan-Meier method, NSCLC patients with high levels of GPC5 expression demonstrated a significantly shorter overall survival time compared with those with low GPC5 expression levels (median postsurgical survival time: 14.0 months vs. 59.0 months, P=0.001). GPC5 expression was also identified as an independent prognostic factor by Cox regression analysis [adjusted hazard ratio: 2.18; 95% confidence interval (CI): 1.35–3.52; P=0.001]. This study suggested that increased levels of GPC5 expression are a poor prognostic marker for NSCLC.

Serum Krebs von den Lungen-6 level as a diagnostic biomarker for interstitial lung disease in Chinese patients

The purpose of this study was to determine the diagnostic and prognostic values of serum KL‐6 levels in Chinese patients with interstitial lung disease (ILDs).

Lesion with morphologic feature of organizing pneumonia (OP) in CT-guided lung biopsy samples for diagnosis of bronchiolitis obliterans organizing pneumonia (BOOP): a retrospective study of 134 cases in a single center

BACKGROUND Small biopsy samples are generally considered inconclusive for bronchiolitis obliterans organizing pneumonia (BOOP) diagnosis despite their potential to reveal organizing pneumonia (OP) pathologically, necessitating risky invasive tissue biopsy during surgery for reliable confirmation. OBJECTIVE OP by CT-guided lung biopsy was to evaluate the role in the diagnosis of BOOP. METHODS A retrospective review of 134 cases with the OP feature in the CT-guided lung biopsy samples between 2004 and 2011 at a single center was conducted. Diagnostic accuracy of OP by CT-guided lung biopsy and clinical-radiographic data alone were compared. RESULTS After exclusion of 11 cases due to pathology with others besides OP and 15 cases for loss to follow-up, 108 were included. Of these, 95 cases and 13 cases were classified as BOOP and non-BOOP group, respectively. Among BOOP group, only 30 were initially diagnosed as BOOP according to the typical clinical and radiographic features. The other 65 cases with atypical features were diagnosed as BOOP mainly based on OP by CT-guided lung biopsy. Among non-BOOP group, one was misdiagnosed as BOOP, and others were not BOOP according to clinical and radiographic findings. Thus, OP by CT-guided lung biopsy produced a diagnostic accuracy of 87.96% (95/108), much higher than 31.25% (30/96) observed using clinical and radiographic data alone. Combined, these techniques produced diagnostic accuracy of 98.96% (95/96). CONCLUSIONS OP by CT-guided lung biopsy can be effectively used as the pathological evidence for BOOP diagnosis and reducing unnecessary surgery.

A Case of Pulmonary Langerhans Cell Histiocytosis with BRAF V600E-negative and MAP2K1-posivtive Mutations Presenting as Diffuse Nodules in Chest High-resolution Computed Tomography After Smoking Cessation

To the Editor: Pulmonary Langerhans cell histiocytosis (PLCH) in adults is a rare disease occurring almost exclusively in smokers. The characteristic high-resolution computed tomography (HRCT) manifestation of PLCH is a combination of cysts (or cavities) and nodules mainly in the upper lung zone.[1] However, not all HRCT patterns of PLCH are typical. Few treatments are effective in current practice regarding PLCH. Targeted therapy with an inhibitor of mutated BRAF (vemurafenib) has been proved effective in Langerhans cell histiocytosis (LCH) harboring BRAF valine at position 600 (V600E) mutation.[2] MAP2K1 mutations are mutually exclusive with BRAF mutations and might have implications for the use of BRAF targeted therapy.[3] Here, we reported a case of PLCH proven by lung biopsy. A 61-year-old man was referred to our institution for evaluation of intermittent cough with white phlegm. He had been a former smoker (cumulative dose = 20 pack/years) and quit approximately one year before. A chest HRCT scan [Figure 1a] revealed, besides emphysema, diffuse nodules up to 1.5 cm in diameter spread in both lung fields without associated lymphadenopathy or pleural effusions. Positron emission tomography-computed tomography (CT) showed diffuse nodules in lung, many of which revealed increased 18F-deoxyglucose uptake (the average standardized uptake value [SUV]: 2.3; maximum SUV: 3.0). Pulmonary function test revealed moderate mixed restriction and obstruction pattern. Routine laboratory tests, ultrasound cardiogram, and bronchoscopy were unremarkable. CT-guided percutaneous transthoracic fine-needle aspiration (FNA) of pulmonary lesions was performed twice, showing atypical epithelial cells. The patient was discharged upon his own request. Figure 1 (a) Chest high-resolution computed tomography findings of the patient. (b) Histopathological findings of the lung biopsy specimen. H and E stain showed cellular nodule with aggregates of Langerhans cells (original magnification ×200); immunohistochemistry ... Two months later, reexamined chest HRCT [Figure 1a] showed a marked increase in the number and size of nodules, with diameter up to 2.0 cm. The patient underwent video-assisted thoracoscopic surgical (VATS) biopsy of the left upper and lower lobes. During the procedure, many palpable nodules were noted in lung parenchyma with diameters ranging from 0.3 to 1.8 cm. A key morphological feature of LCH cells on microscopic examination was noted, which was their highly convoluted nuclear membranes, and the cells strongly expressed Cluster of Differentiation (CD)1a and S100 [Figure 1b]. PLCH diagnosis was made. Treatment started with prednisone (PDN) (0.5 mg·kg−1·d−1 for 1 month). Unfortunately, chest HRCT obtained one month later showed an increase in the number and size of nodular lesions [Figure 1a]. Based on the results of LCH-III,[4] a randomized international clinical trial embarked by the Histiocyte Society, an adjusted chemotherapeutic regimen of methotrexate (MTX) + vindesine (VDS) + PDN was pursued, i.e., MTX 500 mg/m2 once every two weeks intravenous (i.v.), VDS 3 mg/m2 once a week i.v., PDN 30 mg/m2 d1-30 orally (afterward weekly reduction). He experienced WHO Grade 1/2 toxicities including hepatotoxicity, nausea, and vomiting. One month later, chest HRCT showed a partial clearing of nodules [Figure 1a], and his cough resolved. To explore whether the patient would benefit from the BRAF inhibitor therapy, BRAF and MAP2K1 mutations were further tested. Deoxyribonucleic acid (DNA) was extracted from formalin-fixed and paraffin-embedded (FFPE) samples of the surgical biopsy after histological detection of histiocyte-rich areas, using QIAmp DNA FFPE tissue kit (QIAGEN, Germany) according to instructions. Exon 2 and exon 3 of MAP2K1 and exon 15 of BRAF were amplified by polymerase chain reaction and sequenced with primers as listed in Table 1. As indicated in Figure 1c, one silent mutation (code “CAG” to “CAA” at position 610) in exon 2 of MAP2K1 was detected. However, there were no mutations in exon 3 of MAP2K1 and exon 15 of BRAF. The experiment was approved by the Ethics Committee of Drum Tower Hospital, and the patient gave written informed consent. Table 1 Polymerase chain reaction and sequencing primers for MAP2K1 (exon 2 and exon 3) and BRAF (exon 15) Several findings of this case were quite unique and rare. First, the chest HRCT manifestation of this patient was diffuse nodules alone throughout the lung, without formation of cysts or cavities. Second, the onset of disease was nearly one year after smoking cessation. Diffuse lung nodules progressed both in number and size despite smoking cessation. Third, corticosteroid monotherapy failed to control disease progression while intensified combination chemotherapy (MTX + VDS + PDN) showed efficacy. Finally, and particularly, BRAF and MAP2K1 of the lung lesions were sequenced. One MAP2K1 mutation was observed while BRAF V600E mutation was not detected. To the best of our knowledge, the report regarding MAP2K1 mutations in single-system PLCH was seldom and a new mutation site was identified (code “CAG” to “CAA” at position 610 in exon 2 of MAP2K1). These findings differentiate this case from the usual case of PLCH seen in clinical practice. PLCH usually has little systemic damage. In this case, we did not find extrapulmonary disease manifestations. In elder patients with pure nodular forms in chest HRCT, it is especially important to exclude malignancy, which is why this patient received twice CT-guided percutaneous transthoracic FNA followed by VATS lung biopsy. It is generally thought that lung bases are spared in PLCH. However, in this patient, nodules extended into the lower lung zones and costophrenic angles were infiltrated, which might associate with the severity of disease. Persisting uncertainty about the pathogenesis of PLCH has limited current treatment alternatives. Smoking cessation has been regarded as a treatment; however, worsening conditions despite cessation was observed in this case, indicating that PLCH might be induced by antigens in cigarette smoke but not terminated by smoke cessation. The role of smoking cessation on disease progress seems indifferent. In this case, corticosteroid monotherapy failed to control the disease. This patient further received chemotherapy (MTX + VDS + PDN) according to LCH-III.[4] HRCT imaging showed improvement one month later. Even though this case is one-system LCH, involving the lung might associate with rapid progression and worse prognosis, thus a more intensive chemotherapeutic regimen might be necessary as in multisystem LCH. The most common activating somatic mutation in BRAF gene is a substitution of glutamic acid for V600E, which has been identified in a part of LCH cases. This mutation results in constitutive activation of the mitogen-activated protein kinase (MAPK) pathway. MAP2K1 encodes the dual-specificity kinase of the MAPK pathway. MAP2K1 mutations have been demonstrated to confer resistance to BRAF inhibitor therapy in LCH and other neoplasms.[5] A mutation in exon 2 of MAP2K1 was positive in this patient. Even though it is a silent mutation, it indicates there might be another mechanism of MAPK pathway activation in BRAF V600E-negative PLCH. There is clearly a necessity for better therapies for PLCH. Financial support and sponsorship This work was supported by the grants from The Natural Science Foundation of Jiangsu Province (No. BK20130089), and the National Natural Science Foundation of China (No. 81501972). Conflicts of interest There are no conflicts of interest.

Primary pulmonary intravascular large B-cell lymphoma: A report of three cases and literature review

The present study aimed to investigate the clinicopathological features of primary intravascular large B-cell lymphoma (IVLBCL) of the lung. The clinical and histopathological data of three patients, and the literature was reviewed. The Ethics Committees of Drum Tower Hospital approved the current study based on the three cases. Fever and respiratory symptoms were the main presenting symptoms. Serum lactate dehydrogenase and C-reactive protein were significantly increased. Diffuse ground glass opacities or nodular consolidations were seen on high resolution computed tomography. Lung biopsy revealed lymphoma cells in the lumen of small blood vessels. Tumor cells expressed cluster of differentiation 20 and melanoma associated antigen (mutated) 1. Primary pulmonary IVLBCL is extremely rare and its prognosis is poor. Full recognition of its clinical character and improvement of the diagnostic awareness may help to reduce missed diagnosis, and facilitate appropriate treatment.