Intraoperative confocal microscopy for quantitative delineation of low-grade glio

术中共聚焦显微镜定量描绘低级别胶质细胞

基本信息

批准号：
8890436
负责人：
Jonathan T.C. Liu
金额：
$ 34.94万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8890436
关键词：
Aminolevulinic Acid Brain Neoplasms Cell Density Cells Clinic Clinical Clinical Research Clinical Trials Complement Confocal Microscopy Country Devices Diffuse Excision Fluorescence Future Glioma Hand Health Histopathology Hospitals Housing Human Image Image-Guided Surgery Imaging Device Imaging Techniques Imaging technology Infiltration Institutes Label Magnetic Resonance Imaging Measures Mechanics Medical Medical center Microscope Microscopic Microscopy Modeling Morbidity - disease rate Neurologic Neuronavigation Normal tissue morphology Operating Rooms Operative Surgical Procedures Optical Biopsy Optics Outcome Patients Performance Phase III Clinical Trials Population Postoperative Period Proliferating Protocols documentation Publications Publishing Quality of life Rattus Reporting Research Personnel Residual Tumors Resolution Safety Scanning Site Speed Sterility Surgical margins System Techniques Technology Time Tissues Translating Translations Tumor Tissue Universities Variant Work base density design experience fluorescence imaging imaging modality improved in vivo imaging instrumentation neoplastic cell neuro-oncology pre-clinical treatment site tumor

项目摘要

DESCRIPTION (provided by applicant): In recent years, fluorescence image-guided surgery (FIGS) with contrast provided by 5-aminolevulinic acid (5- ALA)-induced PpIX fluorescence has been demonstrated to improve surgical outcomes for glioma patients. However, there are several limitations to these wide-field imaging methods. For example, it is difficult to accurately determine a surgical margin based on subtle variations in fluorescence intensity in glioma tissues, which are diffuse and lack a distinct transition between tumor and normal. Furthermore, wide-field (low- resolution) approaches, such as FIGS and MRI, provide pixel intensities that represent an average value from many cells, resulting in a diminished ability to detect the sparse tumor cell populations at the margins of diffuse gliomas. This problem is exacerbated in low-grade gliomas, where 5-ALA-induced PpIX fluorescence is only generated in rare proliferating cell populations and is undetectable via wide-field FIGS. Recently, Dr. Nader Sanai, a Co-Investigator on this project, has shown that intraoperative cellular-resolution confocal microscopy can be used to visualize the sparse fluorescent cells in low-grade glioma patients treated with 5-ALA. The implications of this finding are highly significant since reported rates of gross-total resection (GTR) have been relatively low for low-grade gliomas (14% to 46%), suggesting a need for improved image-guidance techniques. Here, we propose to optimize a hand-held intraoperative confocal microscope, previously developed by the PI, to visualize low-grade glioma cells that express 5-ALA-induced PpIX, and to demonstrate that this real-time quantitative in vivo imaging technique correlates with MRI and invasive histopathology for margin assessment. This project brings together a proven team of collaborators, including Co-Investigator Olav Solgaard at Stanford University, to develop an optimized optical-sectioning microscope that incorporates a highly robust MicroElectroMechanical System (MEMS) scanner and an achromatic (wavelength-agile) optical design recently published by the PI's lab. Preclinical ex vivo and in vivo imaging will be performed with a highly realistic and infiltrative at model of glioma to demonstrate the ability to correlate the density of cells expressing PpIX fluorescence vs. actual tumor-cell density and histological grade. Clinical studies will leverage the existing expertise of Co-Investigator Nader Sanai, at Barrow Neurological Institute (BNI), with both wide-field FIGS and intraoperative confocal microscopy of PpIX fluorescence in glioma patients. Based on established and approved protocols at BNI, an exploratory clinical study will be performed to correlate the density of PpIX-labeled cells with histopathology, pre-operative MRI (intraoperative neuronavigation), and post- operative MRI. The results of these preliminary studies will justify future clinical investigations to assess patient outcomes such as volumetric extent of resection, overall survival, and neurological morbidity.

描述（由申请人提供）：近年来，已经证明，荧光图像引导的手术（图2）证明了5-氨基乙酸（5- ALA）诱导的PPIX荧光提供的对比度可改善胶质瘤患者的手术结果。但是，这些广阔的成像方法有几个局限性。例如，很难准确根据胶质瘤组织中荧光强度的细微变化来确定手术边缘，这些变化是弥漫性的，缺乏肿瘤和正常之间的明显转变。此外，宽场（低分辨率）方法（例如无花果和MRI）提供了代表许多细胞的平均值的像素强度，从而导致检测到弥漫性神经胶质瘤缘处稀疏的肿瘤细胞群体的能力下降。该问题在低级神经胶质瘤中加剧了，其中5-ALA诱导的PPIX荧光仅在罕见的增殖细胞群体中产生，并且无法通过广阔的无花果发现。最近，该项目的共同投资者Nader Sanai博士表明，术中细胞分辨率共聚焦显微镜可用于可视化接受5-ALA治疗的低级神经胶质瘤患者中稀疏的荧光细胞。这一发现的含义非常重要，因为报道了对于低级神经胶质瘤（14％至46％），总属性切除术（GTR）相对较低，这表明需要改进的图像指导技术。在这里，我们建议优化先前由PI开发的手持术内共共聚焦显微镜，以可视化表达5-ALA诱导PPIX的低级胶质瘤细胞，并证明这种实时定量的体内成像技术与MRI和Invasive Historaporlogy for Margin评估相关。该项目汇集了一支久经考验的合作者团队，包括斯坦福大学的共同投资者Olav Solgaard，开发了优化的光学截面显微镜，其中包含了PIE的实验室最近发布的高度可靠的微电力系统（MEMS）扫描仪和甲状化（Wavelength-Agagile）光学设计。临床前体内和体内成像将在神经胶质瘤模型上具有高度逼真的浸润性，以证明与表达PPIX荧光与实际肿瘤细胞密度和组织学等级的细胞密度相关的能力。临床研究将利用Barrow神经学研究所（BNI）的共同投资者Nader Sanai的现有专业知识，在胶质瘤患者中，PPIX荧光的宽大无花果和术中共聚焦显微镜。基于BNI的已建立和批准方案，将进行一项探索性临床研究，以将PPIX标记细胞的密度与组织病理学，术前MRI（术中神经导航）和手术后MRI相关联。这些初步研究的结果将证明未来的临床研究是合理的，以评估患者结局，例如体积的切除程度，整体生存和神经系统发病率。