Histopathologic and Immunohistochemical Features of Tissue Adherent to the Electr

描述(申请人提供)：射频消融术(RFA)用于治疗不能切除的结肠癌肝转移(CRHM)。该技术旨在造成比靶肿瘤更大的凝固性坏死(CN)，以便创造5-10毫米的边缘，以减少局部肿瘤进展(LTP)并改善预后。在以往的研究中，我们利用细胞增殖标记物Ki67和细胞凋亡标记物Cave Caspase-3的抗体，用组织病理学和免疫组织化学(IHC)方法检测了肝脏恶性肿瘤射频消融术后组织的黏附情况。我们先前的研究表明，当附着在电极上的组织Ki67阳性时，观察到较高的LTP率和较短的进展时间(TTP)。为了确定经形态苏木精-伊红(H&E)染色鉴定的肿瘤细胞是否仍有活力和增殖能力，或者它们是否已经受损，从而表达早期的凋亡标志，用活力和增殖标志物来评估消融肿瘤是非常重要的。这项研究的目的是前瞻性地验证我们的初步结果，并证明从切除的肿瘤组织中获得的组织病理学和免疫组织化学检查可以作为CRHM RFA术后预后的生物标志物。我们的中心假设是，存在活的肿瘤细胞(Mitotracker(MT)Red或OxPhos抗体(AB)和Ki67阳性的肿瘤细胞)会增加不完全消融的可能性。因此，可以预期更高的LTP速率和更短的TTP。为了验证我们的中心假设，我们提出了以下三个具体目标：1.建立在消融肿瘤组织中发现的存活肿瘤(Ox PHOS AB、MT Red和Ki67阳性肿瘤细胞)(附着在电极上并通过针刺活检获得)是CRHM RFA后LTP和治疗失败的独立预测因子。2.使用RFA后动态CT成像，计算肿瘤的灌注和坏死体积，并将其与肿瘤存活的存在(MT Red、OxPhos AB和Ki67阳性的肿瘤细胞)或消融肿瘤组织的凝固性坏死(附着在电极上并通过针吸活检获得)相关联。3.肿瘤的存活情况(OxPhos AB、MT Red和Ki67阳性肿瘤细胞)或消融肿瘤组织凝固性坏死(附在电极上并经针吸活检获得)与外周血中癌胚抗原(CEA)水平的相关性。根据NCI的估计，2008年美国将有10万名新患者被诊断为结肠癌，近5万人将死于结肠癌和直肠癌。高达50%的结肠癌患者会发生肝转移(CRHM)。这些患者的死亡率最高。RFA是一种新的癌症非手术治疗方法，已成功应用于CRHM的治疗。治疗包括用一种特殊的针头烧毁癌症。不幸的是，没有可用的方法来确认在治疗结束时没有残留癌症。我们的项目检查在射频消融电极上发现的组织或用活检针从切除的肿瘤中获取的组织，以确定治疗后是否仍有存活的癌症。组织病理学和免疫组织化学检测是一种新的、微创、安全、简便的检测方法，可作为判断肝脏RFA术后预后的生物标志物。这种组织检查可能允许修改治疗方法，包括重复RFA，这可能会改善CRHM患者的临床结果。组织病理学和免疫组织化学结果也将与RFA后成像相关联。这可能会识别和验证特定的影像表现，可以作为RFA术后结果的替代标记。使用生物显微镜测试和成像技术来衡量干预措施的影响并改进治疗以改善结果是NCI的优先事项。这一信息对大量CRHM患者的治疗至关重要，并可能影响接受RFA治疗的癌症患者的总体人数。与公共卫生相关：美国每年有超过10万人被诊断出患有肝癌，2008年美国将有近5万人死于结肠癌和直肠癌。射频消融(RFA)是一种新的非手术治疗癌症的方法，已成功地应用于肝癌的治疗。治疗包括用一种特殊的针头烧毁癌症。不幸的是，目前还没有可用的方法来确认在治疗结束时没有留下残留的癌症。我们的项目检查结肠癌烧伤后针头上发现的涉及肝脏的组织，以确定治疗结束时是否还有剩余的活体癌症。这一信息可能被用来让那些针头上有癌症证据的患者退缩。这在治疗大量肝癌患者中至关重要，并可能影响接受RFA治疗的癌症患者的总人数。

DESCRIPTION (provided by applicant): Radiofrequency ablation (RFA) is used for the treatment of non-resectable colon cancer hepatic metastases (CRHM). The technique is designed to cause coagulation necrosis (CN) larger than the target tumor in order to create a 5-10 mm margin to diminish local tumor progression (LTP) and improve outcome. In prior studies we showed that tissue adherent to the electrode after RFA of liver malignancies can be examined by histopathology and immunohistochemistry (IHC) using antibodies to Ki67, a marker of cellular proliferation and Cleave Caspase-3, an apoptotic marker (indicative of CN). Our prior studies demonstrated that when tissue adherent to the electrode was positive for Ki67 a higher LTP rate and shorter time to progression (TTP) was observed. The evaluation of ablated tumor with viability and proliferation markers is extremely important in order to determine whether tumor cells identified on morphologic Hematoxylin & Eosin (H & E) stains are still viable and able to proliferate or if they have been damaged so that they express early apoptotic markers. The goal of this study is to prospectively validate our preliminary results and prove that histopathologic and Immunohistochemical examination of tissue obtained from the ablated tumor can be used as a biomarker of outcome after RFA of CRHM. Our central hypothesis is that the presence of viable tumor cells (Mitotracker (MT) Red or OxPhos antibody (AB) and Ki67 positive tumor cells) increases the probability of incomplete ablation. As a result higher LTP rate and shorter TTP can be expected. To test our central hypothesis we propose the following 3 specific aims: 1. Establish that viable tumor (Ox Phos AB, MT Red and Ki67 positive tumor cells) identified in tissue from the ablated tumor (adherent to the electrode and obtained with needle biopsy) is an independent predictor of LTP and treatment failure after RFA of CRHM. 2. Calculate and Correlate the volume of tumor perfusion and necrosis, using post-RFA dynamic CT imaging, with the presence of viable tumor (MT Red, OxPhos AB and Ki67 positive tumor cells) or coagulation necrosis of the tissue from the ablated tumor (adherent to the electrode and obtained with needle biopsy). 3. Correlate the presence of viable tumor (OxPhos AB, MT Red and Ki67 positive tumor cells) or coagulation necrosis of tissue from the ablated tumor (adherent to the electrode and obtained with needle biopsy) with peripheral blood levels of carcinoembryonic antigen (CEA). According to NCI estimations 100,000 new patients will be diagnosed with colon cancer and almost 50,000 will die from colon and rectal cancer in the US in 2008. As many as 50% of patients with colon cancer, develop hepatic metastases (CRHM). These patients have the highest mortality rate. RFA is a new non-surgical therapy for cancer that has been used with success in the treatment of CRHM. The treatment consists of burning the cancer with a special needle. Unfortunately there is no available method to confirm that at the end of the treatment there is no residual cancer left behind. Our project examines tissue that is found on the RFA electrode or obtained with a biopsy needle from the ablated tumor to determine if there is remaining viable cancer after treatment. Histopathologic and immunohistochemical evaluation of tissue adherent to the electrode or obtained from the ablated tumor by needle biopsy is a novel, minimally invasive, safe and simple test that can be used as a prognostic biomarker of outcome after hepatic RFA. This tissue examination may allow treatment modifications, including repeat RFA that may improve clinical outcome for patients with CRHM. The histopathologic and immunohistochemical findings will also be correlated with post RFA imaging. This may identify and validate specific imaging findings that might be used as surrogate markers of outcome after RFA. The use of biospecimen tests and imaging techniques to measure the impact of interventions and refine treatment to improve outcomes is a priority of the NCI. This information is vital in the treatment of a large population with CRHM and may impact the overall population of cancer patients treated with RFA. PUBLIC HEALTH RELEVANCE: Over a100,000 Americans are diagnosed with liver cancer each year and almost 50,000 will die from colon and rectal cancer in the US in 2008. Radiofrequency Ablation (RFA) is a new non-surgical treatment of cancer that has been used with success in the treatment of liver cancers. The treatment consists of burning the cancer with a special needle. Unfortunately there is no method available to confirm that at the end of the treatment there is no residual cancer left behind. Our project examines tissue that is found on the needle after burning of colon cancer that involves the liver to determine if there is remaining live cancer at the end of the treatment. This information may be used to retreat those patients with evidence of cancer on the needle. This is vital in the treatment of a large population with liver cancers and may impact the overall population of cancer patients treated with RFA.