Effect of Cetuximab and EGFR Small Interfering RNA Combination Treatment in NSCLC Cell Lines with Wild Type EGFR and Use of KRAS as a Possible Biomarker for Treatment Responsiveness

[Background] The epidermal growth factor receptor (EGFR) is a therapeutic target for patients with non-small cell lung cancer (NSCLC). Cetuximab is an anti-EGFR monoclonal antibody that inhibits EGFR signaling and proliferation of colorectal cancer and head and neck cancers. Since only few NSCLC patients benefit from cetuximab therapy, we evaluated a novel combination treatment using cetuximab and EGFR small interfering RNA (siRNA) to strongly suppress EGFR signaling and searched for a biomarker in NSCLC cell lines harboring wild-type EGFR. [Methods] Alterations in EGFR and its downstream genes in five NSCLC cell lines (A549, Lu99, 86-2, Sq19 and Ma10) were assessed through sequencing. The protein expression levels of these molecules were assessed through western blotting. The effect of combination treatment was determined through cell proliferation assay, caspase-3/7 assay, invasion assay, and migration assay. [Results] All cell lines were harboring wild-type EGFR, whereas KRAS, PTEN, TP53 and TP53 were mutated in A549 and Lu99; Lu99 and Sq19; Lu99, 86-2, Sq19 and Ma10; and A549, 86-2, and Sq19 cell lines, respectively. PTEN was not expressed in Sq19, and LKB1 was not expressed in both A549 and Sq19. TP53 was not expressed in both A549 and Lu99. The combination of cetuximab and EGFR siRNA significantly suppressed cell proliferation in 86-2, Sq19 and Ma10, which express wild-type KRAS. It induced apoptosis in A549, 86-2 and Ma10 cells, which express wild type PTEN. The combination treatment had no effect either on cell invasion nor migration in all cell lines. [Conclusion] EGFR targeted therapy using the combination of cetuximab and EGFR siRNA is effective in NSCLC cell lines harboring wild-type EGFR. Wild-type KRAS may act as a potential biomarker for response to combination treatment by the induction of apoptosis in cells with wild-type PTEN

Leukocytapheresis for the treatment of acute exacerbation of idiopathic interstitial pneumonias : a pilot study

Objective : Idiopathic interstitial pneumonias (IIPs) are a group of heterogeneous diffuse parenchymal lung disorders of unknown etiology. An acute exacerbation (AE) is an acute respiratory deterioration that occurs in IIPs. The prognosis of AE of IIPs (AE-IIPs) is extremely severe ; however, no established therapies exist. We aimed to evaluate the efficacy of leukocytapheresis (LCAP) to treat patients with AE-IIPs. Patients and Methods : Six chronic IIPs patients who developed AE were enrolled in this study. We performed LCAP on days 2, 3, 9 and 10 in all six patients. All patients were also treated with high-dose corticosteroids and a continuous administration of low-molecular-weight heparin. We observed 30-day survival after the diagnosis of AE to evaluate the efficacy of LCAP. We also assessed oxygenation, high-resolution computed tomography (HRCT) findings, and certain chemical mediators in the peripheral blood. Results : Five of six patients survived more than 30 days. One patient died of progressive respiratory failure. Oxygenation and HRCT findings tended to improve in all survivors. The serum levels of lactate dehydrogenase, high mobility group box-1, and interleukin-18 were significantly decreased statistically post-LCAP. No severe adverse events occurred. Conclusion :We suggest that LCAP is a safe and effective therapy for treating patients with AE-IIPs

Human brain activities relating to mouth movements perception

　脳磁針を用い、「口の動き」がヒトの脳内でどのように情報処理されているかを調べるために以下の二つの実験を行った。(1) 「口の動き」によって誘発される脳活動の時間的、空間的特徴(2) 音声を聞いているときに同時に「口の動き」を提示した際の聴覚野活動に対する影響　まず(1)に関して、仮現運動刺激を利用し「目の動き」をコントロールにして、「口が開く」、「口が閉じる」動きによって誘発される脳活動の違いを調べた。「口の動き」によって明瞭な成分(潜時は約160ミリ秒)が誘発された。次に複数双極子モデル、the brain electic source analysis(BESA)を用いて以下の4つの活動源を推定した。1　後頭側頭部(ヒトのMT/V5野) 2&3左右の第一次視覚野4　側頭葉下面(紡錘状回)　双極子1の活動は他のものに比べて大変大きいものであった。「口の動き」の認知には主として後頭側頭部が関与していることが示された。 「口が開く」と「口が閉じる」という条件より推定された双極子1はほぼ同じ所に位置していた。(1)の実験結果より、「口の動き」、「目の動き」によってヒトのMT/V5野が活動することが示された。また「目の動き」による活動は「口の動き」によるものより大きく、「口が開く」動きと「口が閉じる」動きによって誘発される脳活動には違いはなかった。MT/V5野は動きの認知に関して上側頭溝と異なった過程を持つことがわかった。　次に(2)の実験では、音声聴取の際、「口の動き」を見たときに聴覚野の活動にどういう影響が生じるかを、聴覚野の反応を示すM100(潜時が約100ミリ秒)の時間的、空間的特徴を調べることで明らかにした。　「音声のみの出力」と「音声と口の動きの同時出力」の際のM100を比べると両条件の間に潜時、振幅に関して有意な差はなかった。次にM100の活動源を単一双極子モデルにて推定したところ、両条件ともにヘシェル回に位置し、なおかつその場所に関して有意な差は認められず、またその活動の大きさにおいても有意な差は認められなかった。　(2)の実験結果より、音声聴取の際のへシェル回の活動に対して、「口の動き」の影響はなく音声の特徴を処理していることが示された