Ashfaque A. Memon

Positron Emission Tomography (PET) Imaging with [11C]-Labeled Erlotinib: A Micro-PET Study on Mice with Lung Tumor Xenografts

Erlotinib (Tarceva) targets the epidermal growth factor receptor (EGFR), which is commonly overexpressed in human cancers, including lung cancer. We show that erlotinib can be labeled with [(11)C] by reacting the normethyl precursor with [(11)C]-methyl iodide. By using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide proliferation assay, two lung cancer cell lines (A549 and NCI358) were shown to be less sensitive to erlotinib compared with the lung cancer cell line HCC827. This correlated with higher expression and activity of the EGFR in HCC827 cells as compared with the less sensitive cell lines. Micro-positron emission tomography (PET) and biodistribution of erlotinib was performed with [(11)C]-erlotinib in nude mice bearing xenografts of A549, NCI358, and HCC827 cells. Dynamic micro-PET showed that HCC827 tumors had the highest [(11)C]-erlotinib uptake and retained the activity significantly longer as compared with A549 and NCI358 tumors. Biodistribution of [(11)C]-erlotinib in the xenograft models of lung cancer showed the highest accumulation in the liver. In mice carrying the sensitive cancer cells, the accumulation of [(11)C]-erlotinib was higher in tumors than in the other organs. In contrast, the drug accumulated to a comparable extent in tumors from the less sensitive cancer cells and the other organs. Uptake of [(11)C]-erlotinib in the tumors was 1.6%, 0.7%, and 3.7% (percentage of injected dose/g), in A549, NCI358, and HCC827 cells, respectively. We show for the first time that [(11)C]-erlotinib identifies erlotinib-sensitive tumors. These results pave the road for studies examining the benefit of [(11)C]-erlotinib PET in patients with lung tumors or other tumors overexpressing EGFR.

Erlotinib Accumulation in Brain Metastases from Non-small Cell Lung Cancer: Visualization by Positron Emission Tomography in a Patient Harboring a Mutation in the Epidermal Growth Factor Receptor

Introduction: Drugs directed toward the epidermal growth factor receptor (EGFR), such as erlotinib (Tarceva®) and gefitinib (Iressa®), are used for the treatment of patients with advanced non-small cell lung cancer (NSCLC), including patients with brain metastases. However, whether erlotinib actually enters into brain metastases has not been adequately elucidated. In this study, we investigated the accumulation of [11C]-erlotinib by positron emission tomography (PET) combined with computed tomography (CT) and magnetic resonance imaging (MRI). Methods: A 32-year-old patient with NSCLC and multiple brain metastases was treated with first-line erlotinib. EGFR mutations were determined by analyzing a fine-needle lung tumor biopsy taken before the treatment. A PET/CT of the brain with [11C]-erlotinib was performed during treatment, and a MRI of the head and a CT of the chest were performed pre- and posttreatment. Results: The primary lung tumor displayed an erlotinib-sensitizing exon 19 deletion in the EGFR gene, and [11C]-erlotinib PET/CT showed accumulation in the brain metastases. Posttreatment MRI and CT demonstrated regression of both brain metastases and primary lung tumor. Conclusion: Our data demonstrated that erlotinib accumulated in brain metastases in a NSCLC patient who responded to the treatment.

Determination of 14 Circulating microRNAs in Swedes and Iraqis with and without Diabetes Mellitus Type 2

Background Recent reports suggest that immigrants from Middle Eastern countries are a high-risk group for type 2 diabetes (T2D) compared with Swedes, and that the pathogenesis of T2D may be ethnicity-specific. Deregulation of microRNA (miRNA) expression has been demonstrated to be associated with T2D but ethnic differences in miRNA have not been investigated. The aim of this study was to explore the ethnic specific expression (Swedish and Iraqi) of a panel of 14 previously identified miRNAs in patients without T2D (including those with prediabetes) and T2D. Methods A total of 152 individuals were included in the study (84 Iraqis and 68 Swedes). Nineteen Iraqis and 14 Swedes were diagnosed with T2D. Expression of the 14 selected miRNAs (miR-15a, miR-20, miR-21, miR-24, miR-29b, miR-126, miR-144, miR-150, miR-197, miR-223, miR-191, miR-320a, miR-486-5p, and miR-28-3p) in plasma samples was measured by real-time PCR. Results In the whole study population, the expression of miR-24 and miR-29b was significantly different between T2D patients and controls after adjustment for age, sex, waist circumference, family history of T2D, and a sedentary lifestyle. Interestingly, when stratifying the study population according to country of birth, we found that higher expression of miR-144 was significantly associated with T2D in Swedes (OR = 2.43, p = 0.035), but not in Iraqis (OR = 0.54, p = 0.169). The interaction test was significant (p = 0.017). Conclusion This study suggests that the association between plasma miR-144 expression and T2D differs between Swedes and Iraqis.

Mindfulness group therapy in primary care patients with depression, anxiety and stress and adjustment disorders: randomised controlled trial

BACKGROUND Individual-based cognitive-behavioural therapy (CBT) is in short supply and expensive. AIMS The aim of this randomised controlled trial (RCT) was to compare mindfulness-based group therapy with treatment as usual (primarily individual-based CBT) in primary care patients with depressive, anxiety or stress and adjustment disorders. METHOD This 8-week RCT (ClinicalTrials.gov ID: NCT01476371) was conducted during spring 2012 at 16 general practices in Southern Sweden. Eligible patients (aged 20-64 years) scored ≥10 on the Patient Health Questionnaire-9, ≥7 on the Hospital Anxiety and Depression Scale or 13-34 on the Montgomery-Åsberg Depression Rating Scale (self-rated version). The power calculations were based on non-inferiority. In total, 215 patients were randomised. Ordinal mixed models were used for the analysis. RESULTS For all scales and in both groups, the scores decreased significantly. There were no significant differences between the mindfulness and control groups. CONCLUSIONS Mindfulness-based group therapy was non-inferior to treatment as usual for patients with depressive, anxiety or stress and adjustment disorders.

Diagnostic potential of plasma microRNA signatures in patients with deep-vein thrombosis

For excluding deep-vein thrombosis (DVT), a negative D-dimer and low clinical probability are used to rule out DVT. Circulating microRNAs (miRNAs) are stably present in the plasma, serum and other body fluids. Their diagnostic function has been investigated in many diseases but not in DVT. The aims of present study were to assess the diagnostic ability of plasma miRNAs in DVT and to examine their correlation with known markers of hypercoagulability, such as D-dimer and APC-PCI complex. Plasma samples were obtained from 238 patients (aged 16-95 years) with suspected DVT included in a prospective multicentre management study (SCORE). We first performed miRNA screening of plasma samples from three plasma pools containing plasma from 12 patients with DVT and three plasma pools containing plasma from 12 patients without DVT using a microRNA Ready-to-use PCR Panel comprising 742 miRNA primer sets. Thirteen miRNAs that differentially expressed were further investigated by quantitative real-time (qRT)-PCR in the entire cohort. The plasma level of miR-424-5p (p=0.01) were significantly higher, whereas the levels of miR-136-5p (p=0.03) were significantly lower in DVT patients compared to patients without DVT. Receiver-operating characteristic curve analysis showed the area under the curve (AUC) values of 0.63 for miR-424-5p and 0.60 for miR-136-5p. The plasma level of miR-424-5p was associated with both D-dimer and APC-PCI complex levels (p<0.0001 and p=0.001, respectively). In conclusions, these findings indicate that certain miRNAs are associated with DVT and markers of hypercoagulability, though their diagnostic abilities are probably too low.

Factor H autoantibodies in patients with antiphospholipid syndrome and thrombosis

Objective. Autoantibodies to complement factor H (FH) are associated with atypical hemolytic uremic syndrome, but can also be detected in patients with rheumatoid arthritis and in patients positive for lupus anticoagulants and thus potentially antiphospholipid syndrome (APS). To our knowledge, no data are available on the association between the presence of FH autoantibodies in APS and clinical manifestations. Methods. We determined FH autoantibody levels using ELISA in 2 cohorts of patients with primary (PAPS) and secondary APS (SAPS) from Serbia and Italy, and an additional cohort including patients with venous thromboembolism (VTE) from Sweden. Results. FH autoantibodies were detected in 13.7% of patients (n = 73) with PAPS and 30.3% of patients (n = 33) with SAPS in the Serbian cohort. FH autoantibody frequency in the Italian cohort was 33.3% (n = 15) and 36% (n = 25) in PAPS and SAPS, respectively. Both FH autoantibody levels and frequencies observed in both APS cohorts were significantly higher than in matched healthy controls (5%). Further, patients with PAPS with venous thrombosis in the Serbian cohort had significantly higher levels of FH autoantibodies. Therefore, we analyzed a dedicated Swedish thrombosis cohort and found that patients with FH autoantibody positivity had higher risk of VTE recurrence (HR 2.0, 95% CI 1.2–3.3, p = 0.011) compared with the reference group of FH autoantibody–negative patients. Conclusion. Overall, the data indicate that in patients with APS and recurrent venous thrombosis, there are increased levels of FH autoantibodies, a finding associated with poor clinical outcome.

The epidermal growth factor family has a dual role in deciding the fate of cancer cells

Expression of the epidermal growth factor (EGF) receptors HER1 and HER2 has been implicated in tumour growth and poor survival, whereas expression of HER3 and HER4 has been associated with improved survival of bladder cancer patients. The balance between the expression of the EGF family members may therefore have a role to play in determining the final outcome in cancer cells. To check this, we examined the effect of HER1 activation and inhibition on the expression of the EGF receptors HER3 and HER4 and ligands – the heregulins (HRGs). RT4 bladder cancer cells were treated with 1nM HB‐EGF (known to induce cell proliferation by activating HER1 receptor) and the mRNA content of the two receptors (HER3 and HER4) and their activating ligands (HRG1‐HRG4) was quantified by real time PCR at indicated time‐points. Expressions of HRG1α and HRG1β increased 8‐fold and 9‐fold, respectively, whereas the expressions of HRG2α (4‐fold), HRG2β (2.5‐fold) and HRG4 (3.5‐fold) decreased. In contrast, inhibition of tyrosine kinase activity of HER1 with 5 μM Iressa (a specific inhibitor of HER1) resulted in an increase in mRNA expression of HRG2α (2.5‐fold) and HRG4 (1.5‐fold). In addition, expression of the receptors HER3 (1.5‐fold) and HER4 (2‐fold) was also increased. In conclusion, we demonstrate that activation of the HER1 receptor suppressed the expression of a specific set of HRGs. A decrease in expression of HRG2 and HRG4 during HB‐EGF treatment supports their role in growth inhibition, whereas an increase in HRG1 expression points to a role as a growth stimulatory member of the EGF family.

Role of family history of venous thromboembolism and thrombophilia as predictors of recurrence: a prospective follow‐up study

Several studies have shown that the family history of venous thromboembolism (FHVTE) is a predictor of first venous thromboembolism (VTE). However its role in recurrent VTE is still controversial.

Apolipoprotein M and the risk of unprovoked recurrent venous thromboembolism.

INTRODUCTION Apolipoprotein M (ApoM) protects against atherosclerosis; however, it is unknown whether it also protects against recurrent venous thromboembolism (VTE). MATERIAL AND METHODS Patients in the Malmö Thrombophilia Study (MATS) were followed post-anticoagulant treatment until the diagnosis of recurrent VTE or the end of the study (mean follow-up 36 months). Among patients with a first episode of unprovoked VTE, we identified 43 patients (9.7%) with recurrent VTE during the follow-up period. Three age- and sex-matched control subjects without recurrent VTE were selected for each case (n = 129). Plasma levels of ApoM were quantified by a sandwich ELISA method. RESULTS Among all patients, the plasma levels (mean ± SD) of ApoM were not significantly different between patients with recurrent (0.70 ± 0.2) and non-recurrent VTE (0.74 ± 0.2), p = 0.2. However, after stratification of data according to gender, male patients with recurrent VTE showed significantly (p=0.02) lower ApoM levels (0.63 ± 0.2) as compared to those with non-recurrent VTE (0.74 ± 0.2). No significant differences in ApoM levels were found between recurrent (0.8 ± 0.2) and non-recurrent VTE (0.75 ± 0.2) in female patients, p = 0.3. Cox-regression analysis showed that the risk of recurrent VTE was 0.98 (95% CI, 0.96-0.99) for each 0.01 μM increase in ApoM level in male patients (p = 0.042), and this risk remained unchanged after adjusting for inherited thrombophilia and body mass index (p = 0.027). ApoM levels were not associated with the risk of recurrent VTE in female patients. CONCLUSION Our results show that levels of ApoM in recurrent VTE may differ according to gender and lower levels of ApoM may predict VTE recurrence in male patients.

The association between cytokines and insulin sensitivity in Iraqi immigrants and native Swedes

Objectives To investigate the associations between cytokines and insulin sensitivity in Swedish residents born in Iraq and Swedish residents born in Sweden. Design Cross-sectional study. Settings Iraqi and Swedish origin residents of Rosengård area of Malmö, aged 45–65 years, were randomly selected from the census register. Participants/methods 194 (Iraqi, n=107; Swedish, n=87) participants agreed to participate in the study. Nineteen participants dropped out (Iraqi, n=11; Swedish, n=8). Participants who had already been diagnosed with type 2 diabetes mellitus (T2DM), those who could not participate in an oral glucose tolerance test and those who had a cold/fever at the time of blood sampling were excluded. In total, serum samples from 135 individuals of Swedish (n=62) and Iraqi (n=73) origin were included. Serum concentrations of a panel of 10 cytokines, comprising interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12 (p70), IL-13, interferon-γ and tumour necrosis factor-α were analysed by Luminex multiplex assay. Results In the whole study population, levels of all tested cytokines were inversely associated with insulin sensitivity index (ISI), independent of age, sex, body mass index (BMI), sedentary lifestyle and family history of T2DM (p ≤ 0.05). Interestingly, stratification of the study population according to country of birth showed a significant inverse association between all tested cytokines and ISI in the Iraqi-born population (p ≤ 0.01). The association was independent of age, sex, BMI, sedentary lifestyle and family history of T2DM. In contrast, with the exception for IL-6 (p=0.05), no other tested cytokine was found to be significantly associated with ISI in the Swedish-born population (p≥0.05). Conclusions Our results show an association between cytokines and ISI in the Iraqi-born population but not in the Swedish-born population.