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，利用5-氨基乙酰丙酸（5-ALA）诱导的PpIX荧光提供的对比度的荧光图像引导手术（FIGS）已被证明可以改善神经胶质瘤患者的手术结果。然而，这些宽视场成像方法存在一些局限性。例如，很难准确地根据神经胶质瘤组织中荧光强度的细微变化确定手术切缘，这些组织是弥漫性的并且在肿瘤和正常之间缺乏明显的过渡。此外，宽视野（低分辨率）方法，例如无花果和MRI，提供代表许多细胞平均值的像素强度，导致检测弥漫性神经胶质瘤边缘稀疏肿瘤细胞群的能力下降。这个问题在低级别神经胶质瘤中更为严重，其中 5-ALA 诱导的 PpIX 荧光仅在罕见的增殖细胞群中产生，并且通过宽视野图无法检测到。最近，该项目的联合研究员 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 治疗的低级别神经胶质瘤患者中稀疏的荧光细胞。这一发现的意义非常重大，因为报告的比率低级别胶质瘤的大体全切除 (GTR) 相对较低（14% 至 46%），表明需要改进的图像引导技术。在这里，我们建议优化 PI 先前开发的手持式术中共聚焦显微镜，以可视化表达 5-ALA 诱导的 PpIX 的低级别胶质瘤细胞，并证明这种实时定量体内成像技术与通过 MRI 和侵入性组织病理学进行边缘评估。该项目汇集了一支经过验证的合作者团队，包括斯坦福大学的联合研究员 Olav Solgaard，最近开发了一款优化的光学切片显微镜，该显微镜结合了高度稳健的微机电系统 (MEMS) 扫描仪和消色差（波长灵活）光学设计由 PI 实验室出版。临床前离体和体内成像将在神经胶质瘤模型中进行高度逼真和浸润性成像，以证明将表达 PpIX 荧光的细胞密度与实际肿瘤细胞密度和组织学分级相关联的能力。临床研究将利用巴罗神经学研究所 (BNI) 联合研究员 Nader Sanai 的现有专业知识，对神经胶质瘤患者进行宽视场图和术中 PpIX 荧光共聚焦显微镜检查。根据 BNI 制定和批准的方案，将进行一项探索性临床研究，将 PpIX 标记细胞的密度与组织病理学、术前 MRI（术中神经导航）和术后 MRI 相关联。这些初步研究的结果将证明未来临床研究的合理性，以评估患者的结果，例如切除体积范围、总体生存率和神经发病率。