Stimulated Raman imaging for label-free histology to guide brain tumor surgery

 描述(申请人提供)：脑肿瘤手术的目标是最大限度地切除肿瘤，而不会造成永久性的神经功能障碍。研究表明，患者的预后很大程度上取决于这两个变量。然而，这是困难的，因为在手术室里，肿瘤组织往往与健康组织难以区分。术前和术中MRI通过神经导航可以指导脑肿瘤手术，但由于术中脑变形和脑移位，不能准确显示肿瘤边缘。作为目前的金标准，H&E染色通常用于术中的初步诊断，但由于组织伪影、劳动强度和时间延迟等限制，它不能常规用于指导切除范围。荧光引导手术已开始被用于指导脑肿瘤手术，但对低级别胶质瘤通常不起作用，因为低级别胶质瘤对延长生存时间和提高生活质量构成了最大的挑战和机会。受激拉曼成像(SRI)已被用于活体组织的快速无标记生物分子定位。最近，我们用SRI开发了一种新的方法来成像核酸以及蛋白质和脂肪。核酸的可视化可以定义核的形态和染色质结构，从而能够对临床相关的细胞形态进行病理评估，提供几乎与H&E染色相同的信息。本翻译研究的目标是建立用于脑肿瘤诊断的SRI无标记组织病理学的方法和实践，并最终将其开发成临床设备，用于在手术室中实时地对新鲜组织进行环境SR成像。如果达到这一目标，该项目将通过提供实时组织诊断来更精确地控制切除的程度和保留神经功能，从而极大地改善目前脑肿瘤手术的临床实践。此外，这种方法可能对所有肿瘤手术和其他临床病理，如细针活检和骨髓活检都有价值。该奖项的候选人法克·卢博士是布里格姆妇女医院的博士后研究员。卢博士在生物光子学和生物医学光学方面拥有丰富的经验和专业知识，专门从事生物医学应用的受激拉曼显微镜和多光子显微镜的系统创新。BWH在人类疾病的基础、临床和转化研究方面处于国际领先地位，并建立了多个研究计划，以促进年轻研究人员的工作和职业生涯发展。BWH还拥有国家图像引导治疗中心(P41)和先进的多模式图像引导操作套件(AMIGO)基础设施，这将为拟议的研究提供巨大支持。卢博士的长期研究目标是开发和应用先进的生物物理和光学技术和设备，以提高对癌症等疾病的理解、诊断、治疗和预防，从而更好地提供医疗保健。他的长期职业目标是成为一名在翻译研究前沿工作的独立调查员。五年获奖期的近期目标是1)建立和评估用于脑肿瘤诊断的无标记组织病理学SRI，2)展示用于脑肿瘤诊断的手术切除新鲜组织的环境成像，用于神经外科手术切除的实时指导，3)开发新型受激拉曼显微镜以并行收集全光谱数据用于无标记组织病理学以及脂质生物标志物筛选，4)开发用于SR图像分析的计算机算法，以实现脑肿瘤的自动诊断，以及5)通过集成该仪器和软件，建立并演示用于指导脑肿瘤手术的临床设备的原型。该设备还可用于其他肿瘤手术，具有很大的商业化潜力。为了进行拟议的研究，除了进一步发展他目前在生物光子学、受激拉曼显微镜和非线性光学显微镜方面的优势外，Lu博士还计划接受更多的教育和培训，以丰富和扩展他在以下四个方面的知识和专长：1)接受进行翻译研究的关键培训；2)丰富他对脑肿瘤生物学和医学的知识和加深了解；3)学习为SR图像分析开发计算机算法的技能；以及4)为撰写拨款申请和独立寻求资金做准备。卢博士将参加从哈佛催化剂和冷泉港实验室提供的12个正式课程。他将每周出席在波士顿大学和哈佛大学举行的研讨会，这是由麻省理工学院计算机科学和人工智能实验室举办的生物医学图像分析系列研讨会。他还将参加每年一到两次的年度会议，介绍他的工作供同行讨论。组织了一支强大的指导团队，为拟议的研究和卢博士的职业发展提供坚实的支持，包括BWH神经外科的Alexandra Golby博士、哈佛大学相干拉曼显微镜的X.Sunney Xie教授、BWH分子癌诊断的Nathalie Agar教授、麻省理工学院计算机图像分析的Polina Golland教授和BWH神经病理学的Sandro Santagata博士。这一职业发展奖将为卢博士提供转型为翻译研究独立研究员所需的培训和技能。

 DESCRIPTION (provided by applicant): The goal of brain tumor surgery is to maximize removal of tumor without causing permanent neurologic deficits. Studies have shown that outcomes for patients depend strongly on these two variables. However, this is difficult because tumor tissue is often indistinguishable from healthy tissue in the operating room. Preoperative and intraoperative MRI through neuronavigation can be used to guide brain tumor surgery, but it is not able to accurately delineate the tumor margin due to the brain deformation and brain shift during surgery. H&E staining as the current gold standard is often performed intraoperatively for preliminary diagnosis, but it is not used to guide extent of resection routinely due to the limitations including tissue artifacts, labor-intensiveness, and time delay. Fluorescence guided surgery is beginning to be used to guided brain tumor surgery, but is generally not useful for low grade gliomas, the tumors which pose the greatest challenge and opportunity to extend survival and improve quality of life. Stimulated Raman imaging (SRI) has been used for rapid label-free biomolecular mapping in live tissue. Recently we have developed a novel approach with SRI to image nucleic acids, together with protein and lipids. Visualization of nucleic acids allows definition of nuclear morphology and chromatin structures, thereby enabling pathologic evaluation of clinically relevant cellular morphology, providing almost equivalent information to H&E staining. The goal of this translational research is to establish the methodology and practice for label-free histopathology with SRI for brain tumor diagnosis, and eventually develop it into a clinical device for ambient SR imaging of fresh tissue in the operating room in real time If the aims are achieved, this project will greatly improve the current clinical practice of brain tumor surgery by providing real time tissue diagnosis for more precise control of the extent of resection and preservation of neurologic function. Furthermore, this approach may be of value for all oncologic surgeries and other clinical pathology such as fine needle biopsies and bone marrow biopsy. The candidate for this award Dr. Fake Lu is a postdoctoral research fellow at Brigham and Women's Hospital. Dr. Lu has extensive experience and expertise in biophotonics and biomedical optics, specialized in system innovations of stimulated Raman microscopy and multiphoton microscopy for biomedical applications. BWH is an international leader in basic, clinical and translational research on human diseases, and has established multiple research programs to promote the work and professional career development of young investigators. BWH is also home to the National Center for Image Guided Therapy (P41) and the Advanced Multi-modality Image Guided operating suite (AMIGO) infrastructure which will greatly support to proposed research. Dr. Lu's long term research goal is to develop and apply advanced biophysical and optical technologies and devices to improve understanding, diagnosis, treatment, and prevention of diseases, such as cancers, for better health care. His long-term career goal is to become an independent investigator working at the frontier of translational research. The immediate objectives for the five-year award period is to 1) establish and evaluate label-free histopathology SRI for brain tumor diagnosis, 2) to demonstrate ambient imaging of surgically removed fresh tissue for diagnosis of brain tumors for real time guidance of neurosurgical resection, 3) to develop a novel stimulated Raman microscopy to collect full spectral data in parallel for label-free histopathology, as well as lipid biomarker screening, 4) t develop a computer algorithm for SR image analysis to realize automatic brain tumor diagnosis, and 5) finally by integrating the instrument and software to build and demonstrate a prototype of a clinical device for guiding neurosurgical resection of brain tumors. This device could also be used for other oncologic surgeries and will have great potential for commercialization. To conduct the proposed research, in addition to further develop his current strengths in biophotonics, stimulated Raman microscopy, and nonlinear optical microscopy, Dr. Lu plans to receive more education and training to enrich and expand his knowledge and expertise in the following four areas: 1) to receive critical training in conducting translational research, 2) to enrich his knowledge and deepen his understanding in brain tumor biology and medicine, 3) to learn skills in developing computer algorithms for SR image analysis, and 4) to prepare for writing grant applications and seeking funding independently. Dr. Lu will participate in 12 formal courses selected from those offered by Harvard Catalyst and Cold Spring Harbor Laboratory. He will attend weekly seminars at BWH and Harvard University, seminar series on biomedical image analysis organized by MIT Computer Science and Artificial Intelligence Laboratory. He will also attend one or two annual conferences per year to present his work for peer discussion. A strong mentoring team was organized to provide solid support to the proposed research and Dr. Lu's career development, including Dr. Alexandra Golby in neurosurgery at BWH, Prof. X. Sunney Xie in coherent Raman microscopy at Harvard, Prof. Nathalie Agar in molecular cancer diagnosis at BWH, Prof. Polina Golland in computer-based image analysis at MIT, and Dr. Sandro Santagata in neuropathology at BWH. This career development award will provide Dr. Lu with the training and skills needed to transition into an independent investigator in translational research.