这项研究是支持的案例综合癌症中心（P29 CA43703）。

1。立体定位射波刀放射咨询<ol><li> 说明电脑刀治疗，电脑刀涉及使用类似的使用，使汽车的机器工业机器人手臂上安装的直线加速器。机器人手臂绕着病人直线加速器，它通过多个处理站“的光束传递的进展，在三维空间。跨平面X-射线治疗期间得到验证病人是在一个正确的治疗位置。由于治疗束流配送不仅限于一个二维平面，这个系统进一步增强的能力，为癌症的目标，同时尽量减少辐射剂量，关键的皮肤和内脏器官的结构，以提供集中的辐射剂量。 </li><li> 讨论电脑刀的风险电脑刀放射治疗可能会导致在可能的鞣制和发红的皮肤，疲劳，频繁恶心或diarrh的。EA，罕见的内脏器官损伤，肌肉，神经，骨损伤，第二个癌症的风险非常小。 </li></ol> 2。电脑刀立体定向基准配售<ol><li> 描述基准的位置，为持续性或复发性妇科癌症治疗的妇女进行手术或CT引导下放置至少有三个单1.6×3毫米的黄金软组织内或附近的癌症目标的基准（约米粒大小）。外科剪辑放置在之前的手术时间是足够的密度不是使用电波基准。手术的剪辑不干扰电波针对由于密度差异。 </li><li> 表明电波目标相对基准定位基准点放置在不同的组织深处，周围的电波目标，必须相隔两厘米以上。基准定位在4至6厘米的TARGET。 </li></ol> 3。电脑刀立体定向放射治疗计划<ol><li> 讨论放射外科治疗计划。基准位置后至少有一个星期 （即，使愈合的发生和基准的运动最小化），患者接受CT引导下SBRT治疗模拟。在我们的节目中，两针局部疏散真空袋骨盆固定使用。 </li><li> 表明病人对治疗表。妇女在治疗射波刀放射外科平坦的桌面上的仰卧治疗位置的定位 。 </li><li> 讨论目标和病人固定。病人可能接受疏散真空袋固定，以减少在辐射剂量交付intrafraction议案。一旦固定，妇女接受非对比连续轴向CT高分辨率成像（1.0毫米层厚，间距1对1，电压为120 kVp的，450毫安）。 </li><li> 利用先进的讨论目标定位。成像CT成像后，妇女接受最好的常规18F-FDG PET / CT显像在同一SBRT治疗位置。另外，妇女可以接受传统的对比盆腔磁共振成像。 </li><li> 描述图像放射外科治疗计划合作登记。为Multiplan 3.5.2治疗计划系统（爱可）逆辐射治疗计划进口的高分辨率CT和18架F-FDG的PET / CT检查的图像。 </li><li> 描述团队的方法，放射外科治疗计划。治疗计划，同时放射肿瘤学家和妇科肿瘤轮廓目标癌症临床靶体积（CTVS）。其他疾病的风险组织轮廓和包含在CTVS。如小肠，直肠，膀胱，肝，肾，肺，双侧股骨近端，阴道，骶神经根附近的正常组织结​​构的轮廓由放射肿瘤学家。 </li><li> 描述辐射剂量的处方。辐射处方剂量的3×800 cGy的= 2400 cGy的（俗称的70％等剂量线）已被选中。在200 cGy的生物等效剂量计算，近6170 cGy的交付与处方假设一个“α/β的”肿瘤比10。从SBRT直线加速器发出的辐射是平行使用的12个固定的圆形准直钨（5至60毫米），或用钨铜合金分割六边形光圈准直仪。有14个正常器官的耐辐射和使用烧蚀立体定向放射治疗期间正常组织约束的剂量体积直方图参数列于表1。 </li></ol> 4。电脑刀立体定向放射治疗提供<ol><li>电脑刀SBRT期间，软组织基准执行跨平面图像验证。（或在某些情况下，刚性解剖的标志性建筑，如颅内占位性病变的头骨和脊柱损伤脊椎）平面交叉射线成像跟踪，并确认在预期的目标定位系统（TLS）子系统的目标位置。 </li><li> 验证的目标定位。基准点进行跟踪，在三到六自由度（即X，Y，Z，俯仰，滚动，和偏航）。自动注册的TLS生成的图像，并从最初的治疗计划CT扫描产生的数字重建的X光片（DRRS）相比。 </li><li> 验证的目标定位。如果转向在任何一个自由的六度（X，Y，Z，俯仰，滚转，偏航）已超出预定的公差，基准的自动登记的结果表明，治疗会自动暂停。病人重新定位，然后发生。 </li><li> 考虑目标运动的目标，移动与呼吸运动是TReated利用同步性呼吸系统跟踪子系统（同步性）。与摄像头的使用在治疗室安装，系统连续跟踪贴1病人的胸部周围背心发光二极管（LED）的使用病人的呼吸模式（即，获得外部位置的数据）。10一起与基准点的位置从跨平面X-射线获得的信息（内部位置数据），同步性系统的建立与外部数据和内部数据的关联模型。这种相关性模型允许机械臂按照任何的目标，而在任何交付和所有治疗梁呼吸道引起的议案。 </li><li>治疗可能需要100-150机械臂交付处理的立场。治疗可能会持续30至90分钟。 </li></ol> 5。代表结果立体定向放射治疗（SBRT）可能会涉及到许多个人的辐射光束（<html>蓝色的载体）汇合对单个或多个密切相关的临床辐射的目标，在图1A所示。一位代表良好的电波规划成果，给出了一个具有高辐射剂量的癌症目标量覆盖和癌症的目标保形交付SBRT治疗。 图1B-D显示131束被用来治疗盆腔复发上一个chemorefractory卵巢癌超过42分钟的目标。规定的80％等剂量线SBRT呈现为三个每日800 cGy的分数，总剂量在2400 cGy的保形指数1.94与临床靶区覆盖率100％。剂量体积直方图的临床目标（红色）和直肠（棕色），膀胱（黄色）的关键结构，小肠（浅蓝色）骶神经（棕褐色）和臀部（橙色） 图2描绘。 <img alt="图1" src="/files/ftp_upload/3793/3793fig1.jpg" /> 图1。TP :/ / www.jove.com/files/ftp_upload/3793/3793fig1large.jpg“目标=”_blank“&gt;点击这里查看大图。 <img alt="图2" src="/files/ftp_upload/3793/3793fig2.jpg" /> 图2。剂量体积直方图的临床目标（红色）和直肠（棕色），膀胱（黄色）的关键结构，小肠（浅蓝色）骶神经（棕褐色）和臀部（橙色）。

<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>

（CAK，JMB会员，RD），没有透露。

令人鼓舞的早期SBRT成果推动了放射外科治疗持续性或复发性妇科癌症的临床研究。7，8，15个数据的问题放射生物学效应和SBRT造成的细胞死亡方式。小规模临床研究表明，烧蚀辐射剂量SBRT提供生产有针对性的疾病控制率超过90％。与常规放疗不同，它已具有挑战性SBRT结合放射增敏和细胞毒性/抑制细胞生长的化疗。更好的癌症通过增加规划处理量的扩张，并通过包括18架F-PET / CT图像的目标提高了临床疗效。虽然这是必须考虑其他的方法提供高精度的剂量升级的辐射，它仍然，SBRT是否可以提供同等疗效低和高剂量率近距离未定。事实上，近距离放射治疗是较常用和Vali月技术在妇科癌症的目标实现的辐射剂量递增。正因为如此，热情和谨慎是适当的在读治疗妇科癌症SBRT数据。

<table border="1"><tbody><tr><td> 设备名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>射波刀系统</td><td>爱可（桑尼维尔，CA） </td><td></td><td></td></tr></tbody></table>

stereotactic radiosurgery for gynecologic cancer

立体定向放射治疗（SBRT）区分本身需要更严格的病人固定，占呼吸运动，复杂的治疗计划，板上成像，烧蚀的辐射剂量和数量减少癌症化疗和常规放疗目标通常难治。陡峭的的SBRT辐射剂量下降小康许可证狭窄的铅笔束治疗领域用于烧蚀辐射浓缩成1至3个疗程的治疗。 治疗医生必须明白，SBRT在一个正常组织损伤和地理肿瘤小姐的机会，更大的危险。双方必须解决的癌症目标的固定和高精度治疗交付。癌症目标已经实现了通过使用索引定制泡沫塑料演员，疏散豆袋，或独立于患者腹部压缩体修复模具固定癌症轻盾的1-3 Intrafraction议案。TS因呼吸现在可以减少病人反应屏气技术，4病人的喉舌主动呼吸协调，呼吸相关计算机断层扫描5，6或影像引导植入一个移动的肿瘤内和周围的跟踪基准。7-9射波刀系统（爱可加利福尼亚州桑尼维尔，]）利用安装在一个工业机器人手臂，准确地由一个摄像头，跟踪浸渍装到一个病人的背心上的发光二极管（LED）的设置如下病人呼吸运动的直线加速器辐射。 10放疗利润大幅减少，可以实现运动跟踪，最终使一个较小的规划目标与亚毫米级精度的照射量。11-13 SBRT治疗癌症的目标是通过融合，紧密平行的光束照射。由此造成的辐射剂量的癌症靶体积直方图有更多的代名词CED辐射“肩”，表示高比例的目标，覆盖面和一个小的高剂量辐射的“尾巴。”因此，在降低在SBRT癌症目标的剂量均匀性为代价来增加目标保形。这可能会影响以后的为SBRT目标的肿瘤控制和风险的器官的正常组织的耐受。由于SBRT尖锐剂量衰减，隐匿逃避烧蚀辐射剂量疾病的可能性发生癌症的目标时，没有充分的认可和应用SBRT剂量利润率不足。临床靶区（CTV）的扩展0.5厘米，在一个更大的计划靶体积（PTV），与增加无故正常组织损伤的目标控制。7,8地理小姐的可能性进一步减少，可能会被纳入实现2 - [18F]氟-2 -脱氧-D-葡萄糖（18F-FDG）正电子发射断层显像（PET）SBRT treatme 8 18F-FDG PET / CT的使用。NT规划仅仅是开始尝试发现新的成像目标妇科癌症的分子签名。

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

Stereotactic Radiosurgery for Gynecologic Cancer

难治性化疗或常规放射治疗立体定向放射治疗（SBRT）涉及形象，引导，辐射烧蚀交付癌症的目标。射波刀机器人武装SBRT系统，利用先进的目标定位，提供了大分割放射剂量能够消毒癌症目标。本文将考虑新SBRT妇科癌症的治疗作用。

妇科肿瘤的立体定向放射外科

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 次观看.  University Hospitals Case Medical Center and Case Western Reserve University School of Medicine. 难治性化疗或常规放射治疗立体定向放射治疗（SBRT）涉及形象，引导，辐射烧蚀交付癌症的目标。射波刀机器人武装SBRT系统，利用先进的目标定位，提供了大分割放射剂量能够消毒癌症目标。本文将考虑新SBRT妇科癌症的治疗作用。

视频: 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.

多光谱实时荧光成像术在妇科肿瘤前哨淋巴结的检测

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 ...

科研

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.

立体定向颅内植入和<em在体内</em生物发光成像的肿瘤异种移植的小鼠模型系统的多形性胶质母细胞瘤

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.

癌症治疗监测的切伦科夫发光成像（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.

生物荧光原位模型胰腺癌进展

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.

在乳腺癌细胞雌激素调节小分子RNA的剖析

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.

在组织变性/再生和癌症的研究利用小鼠诱导端粒酶等位基因

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.

研究胰腺癌干细胞特性开发新的治疗策略

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.

动态肺肿瘤跟踪烧蚀立体定向放射身体治疗

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.

与人体组织转化为癌症研究工作

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.

造型卵巢癌传播在人类腹腔的器官文化的早期步骤

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