本研究では、ケース総合がんセンター（P30 CA43703）によってサポートされていました。

1。定位サイバーナイフ放射線外科相談<ol><li> サイバーナイフ放射線治療を説明しています。サイバーナイフ放射線外科は、自動車を作るために使用されるマシンに似て、産業ロボットアームに取り付けられた線形加速器の使用を含む。それはビームデリバリー用に複数の治療法 &quot;の停止&quot;を通じて進行に応じてロボットアームは、患者の周りに三次元空間で線形加速器に移動します。 X線は患者を確認するには、治療中に得られたクロスプレーンは、適切な治療位置にある。治療ビームの配信は2次元平面に限定されるものではないので、このシステムは、さらに重大な皮膚や内臓器官の構造体への放射線量を最小限に抑えながら、がんのターゲットに濃縮された放射線量を提供する能力を向上させます。 </li><li> サイバーナイフの放射線リスクを説明します。サイバーナイフ放射線手術が可能な、皮膚、疲労感、まれに吐き気やdiarrhのなめしと発赤になることがありEA、珍しい内臓損傷、筋肉、神経、骨の損傷、および二次癌の非常に小さなリスク。 </li></ol> 2。定位サイバーナイフ放射線基準点の配置<ol><li> 基準配置を記述します。持続性または再発性婦人科癌の治療を受けている女性は、がんのターゲット内またはその付近に少なくとも3つのシングル1.6×3 mmの金の軟組織のデューシャル（米粒ほどの大きさ）の手術やCTガイドの配置を受けています。手術前の時点で配置され手術クリップのradiosurgicalデューシャルとして使用するのに十分な密度のものではありません。外科用クリップはのradiosurgicalが原因密度の不一致の標的に干渉することはありません。 </li><li> のradiosurgicalターゲットへの相対的な基準位置を示しています。基準点は、のradiosurgicalターゲットの周囲組織の深さを変えるに配置され、2つのセンチ以上で区切らなければなりません。フィデューシャルは、tの4〜6センチ以内に配置されarget。 </li></ol> 3。定位サイバーナイフ放射線治療計画<ol><li> のradiosurgical治療計画を説明します。基準配置後、 少なくとも一週間は （すなわちので、治癒が発生し、基準の動きが最小化されている）、患者はCTガイド下SBRT治療のシミュレーションを受けています。我々のプログラムでは、2つのピンのローカライズされた真空排気バッグ骨盤固定が使用されます。 </li><li> 治療テーブルの上に患者の位置決めを示しています。女性はサイバーナイフのradiosurgical平らなテーブルの上仰臥治療位置に扱われます。 </li><li> ターゲットと患者の固定化について説明します。患者は、放射線の線量配信中intrafractionの動きを減らすために避難させ、真空袋の固定化を受ける可能性があります。一度固定され、女性は非対照的に、連続した軸方向のCT高分解能のイメージングを（1から1ピッチ、1.0mmのスライス厚、電圧120 KVP、450 MAS）を受ける。 </li><li> 高度の利用を議論するターゲットの局在のイメージング。CT撮影後、女性は同じSBRT治療の位置であることが好ましい 、従来の18 F-FDG PET / CTイメージングを受けています。また、女性は、従来の対照的な骨盤の磁気共鳴イメージングを受ける可能性があります。 </li><li> 画像のradiosurgical治療計画の共同登録を説明します。高分解能CTと18 F-FDG PET / CTの画像は、Multiplanの3.5.2治療計画システム（アキュレイ）上の逆の放射線治療計画のためにインポートされます。 </li><li> のradiosurgical治療計画へのチームアプローチについて説明します。治療計画については、放射線腫瘍医と婦人科腫瘍輪郭癌ターゲット臨床標的体積（CTVs）の両方。他の疾患リスクのある組織は、輪郭をされており、CTVsに含まれています。このような小腸、直腸、膀胱、肝臓、腎臓、肺、両側大腿骨近位部、膣、および仙骨神経根などの近くの正常組織構造はで輪郭されている放射線腫瘍医。 </li><li> 線量処方を説明します。3放射線の処方線量×800 cGyで= 2400 cGyでは（一般に70％等線量線に）選択されています。 200 cGyで生物学的等価線量の計算には、ほぼ6170 cGyでは、この処方は、腫瘍のための10の &quot;β/α&quot;の比率を仮定して配信されます。 SBRT線形加速器から放出される放射線は、12の固定タングステンの円形コリメータ（5〜60 mm）のいずれかを使用して、またはタングステン-銅合金セグメント化された六角形IRISコリメータのいずれかを使用してコリメートされる14ノーマル臓器が放射線の耐性を持っており、使用アブレーション定位的放射線治療時の正常組織の制約のために線量体積ヒストグラムパラメータを表1に記載されています。 </li></ol> 4。定位サイバーナイフ放射線の治療配信<ol><li> クロスプレーン画像の検証を実行します。サイバーナイフSBRT時には、軟部組織のフィデュー（例えば、頭蓋内病変の頭蓋骨と脊椎病変の背骨として、またはいくつかのケースでは厳格な解剖学的ランドマーク）は、クロスプレーンX線イメージングにより追跡し、システム（TLS）サブシステムをローカライズするターゲットによって期待されるターゲットの場所にあることが確認されています。 </li><li> ターゲットの位置を確認します。基準点は3つの自由と最大6度（すなわち、X、Y、Z、ピッチ、ロール、ヨー）で追跡されます。 TLSによって生成された画像は自動的に登録され、CTスキャンを計画して初期治療から生成されたディジタル再構成X線写真（DRRs）と比較されます。 </li><li> ターゲットの位置を確認します。自動登録の結果は、フィデューシャルは、6自由度（X、Y、Z、ピッチ、ロール、ヨー）のいずれかで定義済みの許容範囲を超えてシフトしていることを示している場合、処理は自動的に一時停止しています。ポジショニング患者はその後に発生します。 </li><li> ターゲットの動きを考慮してください。呼吸運動とともに移動ターゲットはtrです。同期呼吸器トラッキングサブシステム（同期）を利用eated。治療室に取り付けられたカメラを使用すると、システムは継続的に患者の胸部周囲ベストに貼付されている発光ダイオード（LED）を用いて患者の呼吸パターン（すなわち、外部の位置データを取得）を追跡します。一緒に10クロスプレーンX線から得られた基準位置情報（内部位置データ）を、同期システムは、外部データと内部データとの相関モデルを構築します。この相関モデルは、ロボットアームは、あらゆる治療ビームの配信中にターゲットのいずれかの呼吸誘導運動を追跡することができます。 </li><li>治療は、ロボットアームの100から150までの成果治療ポジションを伴う可能性があります。治療は30〜90分かかる場合があります。 </li></ol> 5。代表的な結果定位体放射線治療（SBRT）は、多くの個々の放射線ビームを含むことができる（<html> 図1Aに示すように、青ベクトル）は、すべて、単一または複数の密接に関連する臨床放射線のターゲットに収束。代表的な良いのradiosurgical計画の成果は、高線量のがん標的体積の範囲と癌ターゲットコンフォーマと成果物SBRT治療を提供します。 図1B-Dのshow 131ビームは42分以上の化学療法抵抗性卵巣癌のターゲットの骨盤再発を治療するために使用された。 SBRTは、80％等線量線は毎日3 800 cGyでの分画に2400 cGyでの総線量のために1.94のコンフォーマリティインデックスと100％臨床標的体積の範囲をレンダリングするために処方した。線量体積臨床標的（赤）および直腸の重要な構造のヒストグラム（茶色）、膀胱（黄色）、小腸（ライトブルー）仙骨神経（日焼け）や腰（オレンジ）は図2に示されている。 <img alt="図1" src="/files/ftp_upload/3793/3793fig1.jpg" /> 図1。TP :/ / www.jove.com/files/ftp_upload/3793/3793fig1large.jpg &quot;ターゲット=&quot; _blank &quot;&gt;拡大図を表示するには、ここをクリックしてください。 <img alt="図2" src="/files/ftp_upload/3793/3793fig2.jpg" /> 図2。臨床標的（赤）および直腸（茶色）、膀胱（黄色）の重要な構造のための線量体積ヒストグラム、小腸（ライトブルー）仙骨神経（日焼け）や腰（オレンジ）。

<ol>
	<li>Sherouse, G., Bourland, J., Reynolds, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virtual+simulation+in+the+clinical+setting:+some+practical+considerations.">Virtual simulation in the clinical setting: some practical considerations.</a> Int. J. Radiat. Oncol. Biol. Phys. 19, 1059-1065 (1990).</li><li>Dickesn, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Personalized+fixation+using+a+vacuum+consolidation+technique.">Personalized fixation using a vacuum consolidation technique.</a> Br. J. Radiol. 54, 257-258 (1981).</li><li>Wunderink, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+respiratory+liver+tumor+motion+by+abdominal+compression+in+stereotactic+body+frame,+analyzed+by+tracking+fiducial+markers+implanted+in+liver.">Reduction of respiratory liver tumor motion by abdominal compression in stereotactic body frame, analyzed by tracking fiducial markers implanted in liver.</a> Int. J. Radiat. Oncol. Biol. Phys. 71, 907-915 (2008).</li><li>Mageras, G., Yorke, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deep+inspiration+breath+hold+and+respiratory+gating+strategies+for+reducing+organ+motion+in+radiation+treatments.">Deep inspiration breath hold and respiratory gating strategies for reducing organ motion in radiation treatments.</a> Semin. Radiat. Oncol. 14, 65-75 (2004).</li><li>Wong, J., Sharpe, M., Jaffray, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+active+breathing+control+(ABC)+to+reduce+margin+for+breathing+motion.">The use of active breathing control (ABC) to reduce margin for breathing motion.</a> Int. J. Radiat. Oncol. Biol. Phys. 44, 911-919 (1999).</li><li>Ford, E., Mageras, G., Yorke, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiration-correlated+spiral+CT:+a+method+of+measuring+respiratory-induced+anatomic+motion+for+radiation+treatment+planning.">Respiration-correlated spiral CT: a method of measuring respiratory-induced anatomic motion for radiation treatment planning.</a> Med. Phys. 30, 88-97 (2003).</li><li>Kunos, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyberknife+radiosurgery+for+squamous+cell+carcinoma+of+the+vulva+after+prior+pelvic+radiation+therapy.">Cyberknife radiosurgery for squamous cell carcinoma of the vulva after prior pelvic radiation therapy.</a> Technol. Cancer Res. Treat. 7, 375-380 (2008).</li><li>Kunos, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stereotactic+body+radiosurgery+for+pelvic+relapse+of+gynecologic+malignancies.">Stereotactic body radiosurgery for pelvic relapse of gynecologic malignancies.</a> Technol. Cancer Res. Treat. 8, 393-400 (2009).</li><li>Kilby, W., Dooley, J., Kuduvalli, G., Sayeh, S., Maurer, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Cyberknife+robotic+radiosurgery+system+in+2010.">The Cyberknife robotic radiosurgery system in 2010.</a> Technol. Cancer Res. Treat. 9, 431-438 (2010).</li><li>Ozhasoglu, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synchrony+&#x2013;+Cyberknife+Respiratory+Compensation+Technology.">Synchrony &#x2013; Cyberknife Respiratory Compensation Technology.</a> Med. Dosimetry. 33, 117-123 (2008).</li><li>Hoogeman, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+Accuracy+of+the+Respiratory+Tumor+Tracking+System+of+the+Cyberknife:+Assessment+By+Analysis+of+Log+Files.">Clinical Accuracy of the Respiratory Tumor Tracking System of the Cyberknife: Assessment By Analysis of Log Files.</a> Int. J. Radiat. Oncol. Biol. Phys. 74, 297-303 (2009).</li><li>Antypas, C., Pantelis, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performance+evaluation+of+a+CyberKnife+G4+image-guided+robotic+stereotactic+radiosurgery+system.">Performance evaluation of a CyberKnife G4 image-guided robotic stereotactic radiosurgery system.</a> Phys. Med. Biol. 53, 4697-4718 (2008).</li><li>Wilcox, E., Daskalov, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+GAFCHROMIC+EBT+film+for+Cyberknife+dosimetry.">Evaluation of GAFCHROMIC EBT film for Cyberknife dosimetry.</a> Med. Phys. 34, 1967-1974 (2007).</li><li>Echner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+design,+physical+properties+and+clinical+utility+of+an+iris+collimator+for+robotic+radiosurgery.">The design, physical properties and clinical utility of an iris collimator for robotic radiosurgery.</a> Phys. Med. Biol. 54, 5359-5380 (2009).</li><li>Choi, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Image-guided+stereotactic+body+radiation+therapy+in+patients+with+isolated+para-aortic+lymph+node+metastases+from+uterine+cervical+and+corpus+cancer.">Image-guided stereotactic body radiation therapy in patients with isolated para-aortic lymph node metastases from uterine cervical and corpus cancer.</a> Int. J. Radiat. Oncol. Biol. Phys. 74, 147-153 (2009).</li></ol>

我々は（CAK、JMB、RD）を開示することは何もない。

励みに早期にSBRT成果は持続性または再発婦人科癌の治療のための放射線治療の臨床研究。7、8を煽っている、15のデータは放射線生物学的効果とSBRTに起因する細胞死のモードに疑問をもっています。小規模臨床研究では、SBRTによって提供されたアブレーション放射線量が90％を超える標的疾患制御率を生成することが示されている。従来の放射線とは異なり、それは放射線増感と細胞増殖/細胞毒性化学療法とSBRTを組み合わせることに挑戦されています。増加した計画処理量の展開を通じて、18 F-FDG PET / CT画像を含めることによって標的優れた癌は臨床転帰を改善しました。それは高精度で用量エスカレート放射線を提供する他の方法を検討することが不可欠ですが、それはSBRTは、低および高線量率小線源治療と同等の治療効果を提供できるかどうかは未定のままです。確かに、小線源治療は、より一般的とワリーです。婦人科がんのターゲットに線量エスカレーションを達成するためのテクニックに日付を記入した。このように、熱意と慎重さの両方が婦人科癌の治療のために利用できるSBRTデータを読み取るのに適しています。

<table border="1"><tbody><tr><td> デバイスの名前 </td><td> 会社 </td><td> カタログ番号 </td><td> コメント </td></tr><tr><td>サイバーナイフシステム</td><td>アキュレイ（サニーベール、カリフォルニア州） </td><td></td><td></td></tr></tbody></table>

stereotactic radiosurgery for gynecologic cancer

定位体放射線治療（SBRT）は、より多くの呼吸運動のために厳格な患者の固定化、会計、複雑な治療計画、オンボードのイメージング、がんの切除放射線量の数を減らし、通常の化学療法と従来の放射線に抵抗性の目標を必要として自身を区別します。 1から3までの治療に凝縮切除、放射線治療に使用される急SBRT線量ドロップオフ許可狭い &quot;ペンシルビーム&quot;治療フィールドに入力します。 治療の医師はそのSBRTは、地理的な腫瘍のミスの正常組織の損傷とチャンスの大きな危機にさらされています感謝しなければなりません。両方が癌ターゲットの固定化により、高精度治療照射によって取り組まれている必要があります。癌のターゲット固定はインデックスカスタマイズされた発泡スチロールのキャスト、避難豆袋、または患者の独立した腹部の圧縮されたボディ修正金型の使用によって達成されています。癌厳しく監督する1-3 Intrafractionの動きは今呼吸に起因するTSは、患者の応答息ホールドのテクニック、4患者のマウスピースアクティブな呼吸の調整、5呼吸相関CT検査、6または移動腫瘍内や周囲に注入フィデューシャルの画像誘導トラッキング削減することができます。7月9日サイバーナイフシステム（アキュレイ[サニーベール、カリフォルニア州]）は、正確に患者に取り付けられたベストを含浸させた発光ダイオード（LED）のセットをカメラが追跡することによって患者の呼吸運動の後に、産業ロボットアームに搭載された放射線線形加速器を利用しています。放射線療法のマージンで10大幅に削減モーショントラッキングによって達成することができる、最終的にはサブミリの精度で照射されるより小さい計画のターゲット·ボリュームをレンダリングします。11月13日 がんはSBRTによって治療目標は、しっかりとコリメートビームを収束して照射される。がん標的体積のヒストグラムに結果放射線量はより多くの代名詞を持っている高い割合の目標カバレッジと小さな高線量の放射線を示すCED放射線 &quot;肩&quot; &quot;テール&quot;したがって、増加のターゲットコンフォーマリティは、SBRT癌ターゲットに減少した線量均一性を犠牲にしています。これはSBRTターゲット内の後続の腫瘍コントロールとリスクのある臓器の正常組織の許容範囲の両方に影響を与える可能性があります。癌のターゲットが完全に認識されず、不十分なSBRT線量マージンが適用されたときにSBRTの急激な線量減衰のために、切除、放射線量をエスケープオカルト疾患の可能性が発生します。大規模な計画標的体積（PTV）の結果0.5センチメートルによる臨床標的体積（CTV）の拡大は、過度の正常組織の損傷せずに増加したターゲットのコントロールに関連付けられている。地理的なミスの確率で7,8さらなる削減が混入することによって達成することができるの2 - [18 F]フルオロ-2 -デオキシ-D-グルコース（18 F-FDG）陽電子放射断層撮影法（PET）SBRT treatmeにおける18 F-FDG PET / CTの8を使用。NTの計画では、婦人科癌のための新しいイメージング標的分子の署名を発見するための試みの始まりに過ぎない。

Watch this Scientific Journal Video about Stereotactic Radiosurgery for Gynecologic Cancer at JoVE.com

Stereotactic Radiosurgery for Gynecologic Cancer

定位体放射線治療（SBRT）は、化学療法または、従来の放射線治療に難治性の癌のターゲットに配信画像誘導、アブレーション放射線が含まれます。洗練されたターゲットの位置推定を用いたロボットの武装サイバーナイフSBRTシステムは、癌のターゲットを殺菌することができるhypofractionated放射線量を提供しています。この資料では、婦人科癌に対するSBRTの新たな治療の役割を検討します。

婦人科癌に対する定位放射線

Stereotactic body radiotherapy (SBRT) distinguishes itself by necessitating more rigid patient immobilization, accounting for respiratory motion, ...

stereotactic-radiosurgery-for-gynecologic-cancer

Research

JoVE Journal

Medicine

17.5K ビュー.  University Hospitals Case Medical Center and Case Western Reserve University School of Medicine. 定位体放射線治療（SBRT）は、化学療法または、従来の放射線治療に難治性の癌のターゲットに配信画像誘導、アブレーション放射線が含まれます。洗練されたターゲットの位置推定を用いたロボットの武装サイバーナイフSBRTシステムは、癌のターゲットを殺菌することができるhypofractionated放射線量を提供しています。この資料では、婦人科癌に対するSB...

ビデオ: Stereotactic Radiosurgery for Gynecologic Cancer

Multispectral Real-time Fluorescence Imaging for Intraoperative Detection of the Sentinel Lymph Node in Gynecologic Oncology

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical excision of the tumor with a full lymphadenectomy in the drainage area of the tumor. However, removal of lymph nodes is associated with increased morbidity due to infection, wound breakdown and lymphedema.4,5 As an alternative, the sentinel lymph node procedure (SLN) was developed several decades ago to detect the first draining lymph node from the tumor.6 In case of lymphogenic dissemination, the SLN is the first lymph node that is affected (Figure 1). Hence, if the SLN does not contain metastases, downstream lymph nodes will also be free from tumor metastases and need not to be removed. The SLN procedure is part of the treatment for many tumor types, like breast cancer and melanoma, but also for cancer of the vulva and cervix.7 The current standard methodology for SLN-detection is by peritumoral injection of radiocolloid one day prior to surgery, and a colored dye intraoperatively. Disadvantages of the procedure in cervical and vulvar cancer are multiple injections in the genital area, leading to increased psychological distress for the patient, and the use of radioactive colloid.
Multispectral fluorescence imaging is an emerging imaging modality that can be applied intraoperatively without the need for injection of radiocolloid. For intraoperative fluorescence imaging, two components are needed: a fluorescent agent and a quantitative optical system for intraoperative imaging. As a fluorophore we have used indocyanine green (ICG). ICG has been used for many decades to assess cardiac function, cerebral perfusion and liver perfusion.8 It is an inert drug with a safe pharmaco-biological profile. When excited at around 750 nm, it emits light in the near-infrared spectrum around 800 nm. A custom-made multispectral fluorescence imaging camera system was used.9.
The aim of this video article is to demonstrate the detection of the SLN using intraoperative fluorescence imaging in patients with cervical and vulvar cancer. Fluorescence imaging is used in conjunction with the standard procedure, consisting of radiocolloid and a blue dye. In the future, intraoperative fluorescence imaging might replace the current method and is also easily transferable to other indications like breast cancer and melanoma.

Fluorescence imaging is a promising innovative modality for image-guided surgery in surgical oncology. In this video we describe the technical procedure for detection of the sentinel lymph node using fluorescence imaging as showcased in gynecologic oncologicy. A multispectral fluorescence camera system, together with the fluorescent agent indocyanine green, is applied.

婦人科腫瘍におけるセンチネルリンパ節の術中検出のためのマルチスペクトルのリアルタイム蛍光イメージング

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical ...

医学

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality treatment consisting of surgical resection, post-operative radiation therapy and temozolomide chemotherapy 1. New therapeutic approaches are clearly needed to improve patient survival and quality of life. The development of more effective treatment strategies would be aided by animal models of GBM that recapitulate human disease yet allow serial imaging to monitor tumor growth and treatment response. In this paper, we describe our technique for the precise stereotactic implantation of bio-imageable GBM cancer cells into the brains of nude mice resulting in tumor xenografts that recapitulate key clinical features of GBM 2. This method yields tumors that are reproducible and are located in precise anatomic locations while allowing in vivo bioluminescent imaging to serially monitor intracranial xenograft growth and response to treatments 3-5. This method is also well-tolerated by the animals with low perioperative morbidity and mortality.

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

We describe an integrated method for the precise, stereotactic implantation of human glioblastoma multiforme cells into the brains of nude mice and subsequent serial in vivo imaging to monitor growth and response to treatment of the resultant xenografts.

定位頭蓋内注入と<emインビボ&gt;</em多形性膠芽腫のマウスモデル系における腫瘍異種移植片の&gt;生物発光イメージング

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality ...

Cerenkov Luminescence Imaging (CLI) for Cancer Therapy Monitoring

In molecular imaging, positron emission tomography (PET) and optical imaging (OI) are two of the most important and thus most widely used modalities1-3. PET is characterized by its excellent sensitivity and quantification ability while OI is notable for non-radiation, relative low cost, short scanning time, high throughput, and wide availability to basic researchers. However, both modalities have their shortcomings as well. PET suffers from poor spatial resolution and high cost, while OI is mostly limited to preclinical applications because of its limited tissue penetration along with prominent scattering optical signals through the thickness of living tissues.
 Recently a bridge between PET and OI has emerged with the discovery of Cerenkov Luminescence Imaging (CLI)4-6. CLI is a new imaging modality that harnesses Cerenkov Radiation (CR) to image radionuclides with OI instruments. Russian Nobel laureate Alekseyevich Cerenkov and his colleagues originally discovered CR in 1934. It is a form of electromagnetic radiation emitted when a charged particle travels at a superluminal speed in a dielectric medium7,8. The charged particle, whether positron or electron, perturbs the electromagnetic field of the medium by displacing the electrons in its atoms. After passing of the disruption photons are emitted as the displaced electrons return to the ground state. For instance, one 18F decay was estimated to produce an average of 3 photons in water5. 
 Since its emergence, CLI has been investigated for its use in a variety of preclinical applications including in vivo tumor imaging, reporter gene imaging, radiotracer development, multimodality imaging, among others4,5,9,10,11. The most important reason why CLI has enjoyed much success so far is that this new technology takes advantage of the low cost and wide availability of OI to image radionuclides, which used to be imaged only by more expensive and less available nuclear imaging modalities such as PET.
 Here, we present the method of using CLI to monitor cancer drug therapy. Our group has recently investigated this new application and validated its feasibility by a proof-of-concept study12. We demonstrated that CLI and PET exhibited excellent correlations across different tumor xenografts and imaging probes. This is consistent with the overarching principle of CR that CLI essentially visualizes the same radionuclides as PET. We selected Bevacizumab (Avastin; Genentech/Roche) as our therapeutic agent because it is a well-known angiogenesis inhibitor13,14. Maturation of this technology in the near future can be envisioned to have a significant impact on preclinical drug development, screening, as well as therapy monitoring of patients receiving treatments.

Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring

Use of Cerenkov Luminescence Imaging (CLI) for monitoring preclinical cancer treatment is described here. This method takes advantage of Cerenkov Radiation (CR) and optical imaging (OI) to visualize radiolabeled probes and thus provides an alternative to PET in preclinical therapeutic monitoring and drug screening.

がん治療のモニタリングのためのチェレンコフ発光イメージング（CLI）

In  molecular imaging, positron emission tomography (PET) and optical imaging (OI)  are two of the most important and thus most widely used modalities1-3. ...

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis.

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

膵臓癌の進行の生物発光同所性モデル

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

Profiling of Estrogen-regulated MicroRNAs in Breast Cancer Cells

Estrogen plays vital roles in mammary gland development and breast cancer progression. It mediates its function by binding to and activating the estrogen receptors (ERs), ER&#945;, and ER&#946;. ER&#945; is frequently upregulated in breast cancer and drives the proliferation of breast cancer cells. The ERs function as transcription factors and regulate gene expression. Whereas ER&#945;&#39;s regulation of protein-coding genes is well established, its regulation of noncoding microRNA (miRNA) is less explored. miRNAs play a major role in the post-transcriptional regulation of genes, inhibiting their translation or degrading their mRNA. miRNAs can function as oncogenes or tumor suppressors&#160;and are also promising biomarkers. Among the miRNA assays available, microarray and quantitative real-time polymerase chain reaction (qPCR) have been extensively used to detect and quantify miRNA levels. To identify miRNAs regulated by estrogen signaling in breast cancer, their expression in ER&#945;-positive breast cancer cell lines were compared before and after estrogen-activation using both the &#181;Paraflo-microfluidic microarrays and Dual Labeled Probes-low density arrays. Results were validated using specific qPCR assays, applying both Cyanine dye-based and Dual Labeled Probes-based chemistry. Furthermore, a time-point assay was used to identify regulations over time. Advantages of the miRNA assay approach used in this study is that it enables a fast screening of mature miRNA regulations in numerous samples, even with limited sample amounts. The layout, including the specific conditions for cell culture and estrogen treatment, biological and technical replicates, and large-scale screening followed by in-depth confirmations using separate techniques, ensures a robust detection of miRNA regulations,&#160;and eliminates false positives and other artifacts. However, mutated or unknown miRNAs, or regulations at the primary and precursor transcript level, will not be detected. The method presented here represents a thorough investigation of estrogen-mediated miRNA regulation.

Molecular signaling through both estrogen and microRNAs are critical in breast cancer development and growth. Estrogen activates the estrogen receptors, which are transcription factors. Many transcription factors can regulate the expression of microRNAs, and estrogen-regulated microRNAs can be profiled using different large-scale techniques.

乳癌細胞におけるエストロゲン調節マイクロRNAのプロファイリング

Estrogen plays vital roles in mammary gland development and breast cancer progression. It mediates its function by binding to and activating the estrogen ...

Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that cause tissue degeneration during ageing. Cancer, with incidence rates greatly increasing with age, is characterized by short telomere lengths and high telomerase activity. To study the roles of telomere dysfunction and telomerase reactivation in ageing and cancer, the protocol shows how to generate two murine inducible telomerase knock-in alleles 4-Hydroxytamoxifen (4-OHT)-inducible TERT-Estrogen Receptor (mTERT-ER) and Lox-Stopper-LoxTERT (LSL-mTERT). The protocol describes the procedures to induce telomere dysfunction and reactivate telomerase activity in mTERT-ER and LSL-mTERT mice in vivo. The representative data show that reactivation of telomerase activity can ameliorate the tissue degenerative phenotypes induced by telomere dysfunction. In order to determine the impact of telomerase reactivation on tumorigenesis, we generated prostate tumor model G4 PB-Cre4 PtenL/L p53L/L LSL-mTERTL/L and thymic T-cell lymphoma model G4 Atm-/- mTERTER/ER. The representative data show that telomerase reactivation in the backdrop of genomic instability induced by telomere dysfunction can greatly enhance tumorigenesis. The protocol also describes the procedures used to isolate neural stem cells (NSCs) from mTERT-ER and LSL-mTERT mice and reactivate telomerase activity in NSCs in vitro. The representative data show that reactivation of telomerase can enhance the self-renewal capability and neurogenesis in vitro. Finally, the protocol describes the procedures for performing telomere FISH (Fluorescence In Situ Hybridization) on both mouse FFPE (Formalin Fixed and Paraffin Embedded) brain tissues and metaphase chromosomes of cultured cells.

Telomere and telomerase play essential roles in ageing and tumorigenesis. The goal of this protocol is to show how to generate two murine inducible telomerase knock-in alleles and how to utilize them in the studies of tissue degeneration/regeneration and cancer.

組織の変性/再生と癌の研究におけるマウス誘導テロメラーゼ対立遺伝子を利用

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors 1,2. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

新しい治療戦略を開発するための膵臓癌幹細胞特性を研究

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Dynamic Lung Tumor Tracking for Stereotactic Ablative Body Radiation Therapy

Physicians considering stereotactic ablative body radiation therapy (SBRT) for the treatment of extracranial cancer targets must be aware of the sizeable risks for normal tissue injury and the hazards of physical tumor miss. A first-of-its-kind SBRT platform achieves high-precision ablative radiation treatment through a combination of versatile real-time imaging solutions and sophisticated tumor tracking capabilities. It uses dual-diagnostic kV x-ray units for stereoscopic open-loop feedback of cancer target intrafraction movement occurring as a consequence of respiratory motions and heartbeat. Image-guided feedback drives a gimbaled radiation accelerator (maximum 15 x 15 cm field size) capable of real-time &#177;4 cm pan-and-tilt action. Robot-driven &#177;60&#176; pivots of an integrated &#177;185&#176; rotational gantry allow for coplanar and non-coplanar accelerator beam set-up angles, ultimately permitting unique treatment degrees of freedom. State-of-the-art software aids real-time six dimensional positioning, ensuring irradiation of cancer targets with sub-millimeter accuracy (0.4 mm at isocenter). Use of these features enables treating physicians to steer radiation dose to cancer tumor targets while simultaneously reducing radiation dose to normal tissues. By adding respiration correlated computed tomography (CT) and 2-[18F] fluoro-2-deoxy-&#7429;-glucose (18F-FDG) positron emission tomography (PET) images into the planning system for enhanced tumor target contouring, the likelihood of physical tumor miss becomes substantially less1. In this article, we describe new radiation plans for the treatment of moving lung tumors.

Stereotactic ablative body radiation therapy (SBRT) involves the precise delivery of high-dose radiation to cancer tumor targets. A novel SBRT platform offers a first-of-its-kind gimbaled radiation accelerator mounted within a pivoting O-ring gantry capable of dynamic image-guided tumor tracking. Here, we describe dynamic tumor tracking for lung targets.

定位アブレーションボディ放射線治療のための動的肺腫瘍の追跡

Physicians considering stereotactic ablative body radiation therapy (SBRT) for the treatment of extracranial cancer targets must be aware of the sizeable ...

Working with Human Tissues for Translational Cancer Research

Medical research for human benefit is greatly impeded by the necessity for human tissues and subjects. However, upon obtaining consent for human specimens, precious samples must be handled with the greatest care in order to ensure integrity of organs, tissues, and cells to the highest degree. Unfortunately, tissue processing by definition requires extraction of tissues from the host, a change which can cause great cellular stress and have major repercussions on subsequent analyses. These stresses could result in the specimen being no longer representative of the site from which it was retrieved. Therefore, a strict protocol must be adhered to while processing these specimens to ensure representativeness. The desired assay(s) must also be taken into consideration in order to ensure that an optimal technique is used for sample processing. Outlined here is a protocol for tissue retrieval, processing and various analyses which may be performed on processed tissue in order to maximize downstream production from limited tissue samples.

Translational cancer research is dependent on extraction of human tissues. Much work has gone into optimizing extraction methods for ex vivo analysis. Here, we describe tissue processing methods allowing for maximal data output from limited samples.

トランスレーショナルがん研究のためのヒト組織での作業

Medical research for human benefit is greatly impeded by the necessity for human tissues and subjects. However, upon obtaining consent for human ...

Modeling the Early Steps of Ovarian Cancer Dissemination in an Organotypic Culture of the Human Peritoneal Cavity

The pattern of ovarian cancer metastasis is markedly different from that of most other epithelial tumors, because it rarely spreads hematogenously. Instead, ovarian cancer cells exfoliated from the primary tumor are carried by peritoneal fluid to metastatic sites within the peritoneal cavity. These sites, most notably the abdominal peritoneum and omentum, are organs covered by a mesothelium-lined surface. To investigate the processes of ovarian cancer dissemination, we assembled a complex three-dimensional culture system that reconstructs the lining of the peritoneal cavity in vitro. Primary human fibroblasts and mesothelial cells were isolated from human omentum. The fibroblasts were then mixed with extracellular matrix and covered with a layer of the primary human mesothelial cells to mimic the peritoneal and omental surfaces encountered by metastasizing ovarian cancer cells. The resulting organotypic model is, as shown, used to examine the early steps of ovarian cancer dissemination, including cancer cell adhesion, invasion, and proliferation. This model has been used in a number of studies to investigate the role of the microenvironment (cellular and acellular) in early ovarian cancer dissemination. It has also been successfully adapted to high throughput screening and used to identify and test inhibitors of ovarian cancer metastasis.

Here, we present a protocol to construct a three-dimensional in vitro model of the lining of the peritoneal cavity, composed of primary human mesothelial cells and fibroblasts layered with extracellular matrix, as a tool to investigate ovarian cancer cell adhesion, invasion, and proliferation.

ヒト腹腔器官培養における卵巣癌普及の初期段階のモデル化

The pattern of ovarian cancer metastasis is markedly different from that of most other epithelial tumors, because it rarely spreads hematogenously. ...