A Wireless Multi-function Microscope for Lifetime Imaging of the Brain Tumor Vasculome

摘要临床前和临床证据表明，脑肿瘤可以改变大脑的结构和功能。 中枢神经系统微血管构筑(即中枢神经系统血管构筑)在进展、治疗和出现 治疗抵抗力。脑肿瘤进展相关的血管重构通过血管生成(即新的血液)发生 舰船编队)。相反，非血管生成途径，如‘共同选择’(即，肿瘤细胞劫持现有血液 血管)和‘免疫调节’(即免疫细胞渗透引起的血管变化)参与 抗血管生成耐药和免疫治疗逃避。为了阐明这些血管生成和非血管生成因子的作用 脑肿瘤进展和治疗反应中的血管生成途径需要影像的发展 可以表征早期到晚期中枢神经系统血管小体变化的工具(即在疾病的整个生命周期内)。 因此，我们的目标是建立一种无线即插即用的多通道显微镜，能够成像结构/功能 在脑肿瘤的整个生命周期中，体内的微血管(~7-10微米)会发生变化。我们计划利用在以下方面的进展 微型化光学、图像传感器设计和无线技术，以制造微型无线显微镜 三个通道：荧光(FL)成像荧光脑瘤细胞或染料；本征光学信号(IO)成像 脑血容量(CBV)和血管内氧合(HbSat)；以及激光散斑对比(LSC)成像 血流量(CBF)。在令人信服的初步数据指导下，我们将追求以下具体目标：(1)制定 具有片上压缩传感和无线传输的无绳系多通道显微镜；(2)表征In 血管生成和共视患者衍生(PDX)脑瘤模型的体内血管组； (3)研究脑肿瘤免疫微环境对体内血管构型的影响。在……下面 目的1我们将制作一种专门用于超低功率无线操作的压缩传感图像传感器。之后 对照同等的台式成像系统进行验证，我们将成像健康小鼠的中枢神经系统血管 麻醉剂的混杂作用。这将包括用FL识别微血管类型(即动脉和静脉)，量化 血管形态和HbSat与IOS、LSC灌注和通过相互关联标测“微血管连通性” 微血管的CBV(或CBF)波动。在AIM2下，我们将描述临床上中枢神经系统血管体的差异 相关的血管生成和共视患者来源的异种移植物，并评估前者是否在 血管重塑导致的微血管连通性。在Aim3下，我们将描述中枢神经系统的活体差异 野生型和非免疫抑制异种移植物的血管小体，以确定是否缓解免疫抑制 增加CBV/CBF/HbSat并促进肿瘤相关巨噬细胞的募集()。我们将创造一个多功能的 3D打印即插即用无线显微镜，允许在任何时间对自由行为的动物进行神经成像 持续时间，在任何任务或生理记录期间(例如脑电)。由于这种显微镜可以为任何荧光团定制， 对光遗传学或药物输送进行改进，并用于行为研究，我们相信它将开启大脑的新纪元 癌症研究，对涉及中枢神经系统血管的疾病(如中风、阿尔茨海默病)具有实用价值。

ABSTRACTPreclinical and clinical evidence has shown that brain tumors can alter the structure and function of the central nervous system microvasculature (i.e. CNS vasculome) during progression, therapy and the emergence of therapeutic resistance. Brain tumor progression related vasculome remodeling occurs via angiogenesis (i.e. new blood vessel formation). In contrast, non-angiogenic pathways such as ‘co-option’ (i.e. tumor cells hijacking extant blood vessels) and ‘immunomodulation’ (i.e. vascular changes induced by the infiltration of immune cells) are involved in antiangiogenic resistance and immunotherapy evasion, respectively. To elucidate the role of these angiogenic and non- angiogenic pathways on brain tumor progression and therapeutic response necessitates the development of imaging tools that can characterize early to advanced in vivo changes in the CNS vasculome (i.e. over the lifetime of the disease). Therefore, our goal is to build a wireless ‘plug-n-play’ multichannel microscope capable of imaging structural/functional microvascular (~7-10 µm) changes in vivo, over the entire lifetime of a brain tumor. We propose to exploit advances in miniaturized optics, image sensor design and wireless technology to fabricate a miniature, wireless microscope with three channels: fluorescence (FL) to image fluorescent brain tumor cells or dyes; intrinsic optical signals (IOS) to image cerebral blood volume (CBV) and intravascular oxygenation (HbSat); and laser speckle contrast (LSC) to image cerebral blood flow (CBF). Guided by compelling preliminary data, we will pursue the following Specific Aims: (1) Develop a tether-free multichannel microscope with on-chip compressed sensing and wireless transmission; (2) Characterize the in vivo vasculome in angiogenic and co-optive patient-derived (PDX) brain tumor models over their lifetime; and (3) Characterize in vivo changes in the vasculome induced by the immune microenvironment of brain tumors. Under Aim1 we will fabricate a specialized image sensor with compressed sensing for ultra-low power wireless operation. After validation against an equivalent benchtop imaging system, we will image the CNS vasculome in healthy mice without the confounding effects of anesthetics. This will include identifying microvessel type (i.e. artery vs. vein) with FL, quantifying vascular morphology and HbSat with IOS, perfusion with LSC, and mapping ‘microvascular connectivity’ by correlating CBV (or CBF) fluctuations in microvessels. Under Aim2 we will characterize differences in the CNS vasculomes of clinically relevant angiogenic and co-optive patient-derived xenografts, and assess if the former exhibits larger disruptions in microvascular connectivity due to vascular remodeling. Under Aim3, we will characterize in vivo differences in the CNS vasculomes of wild-type and non-immunosuppressed xenografts, to determine if alleviating immunosuppression increases CBV/CBF/HbSat and promotes recruitment of tumor associated macrophages (TAM). We will create a versatile 3D printed plug-n-play wireless microscope that permits neuroimaging in freely behaving animals at any time, for any duration, during any task or physiological recording (e.g. EEG). As this microscope can be customized to any fluorophore, modified for optogenetics or drug delivery, and used for behavioral studies, we believe it will usher in a new era of brain cancer research, with utility in diseases involving the CNS vasculome (e.g. stroke, Alzheimer’s disease).