Molecular Imaging of the Extracellular Matrix and its Avenues for Metastasis

在研究部分4中，我们将开发和临床测试内窥镜I型胶原（科尔）纤维 使用创新的超紧凑型光纤活检针兼容显微内镜进行成像， 优化用于科尔纤维的二次谐波发生（SHG）显微检测。我们将把这个 临床前SHG科尔成像的常规临床14号活检针内镜 和临床研究。活检时将在原发性乳腺肿瘤中检测到科尔纤维，以预测 乳腺癌患者的淋巴结转移。淋巴结受累的评估， 在乳腺癌患者的早期临床管理中， 在最初乳腺活检的早期选择治疗方案具有显著优势。 科尔纤维是乳腺肿瘤细胞外基质（ECM）的重要组成部分， 基质科尔纤维促进乳腺肿瘤形成、侵袭和转移。高科尔纤维密度， 原发性肿瘤促进转移，因为癌细胞在肿瘤内沿沿着致密的CoH纤维途径迁移， 转移性原发性肿瘤形成转移性结节。因此，我们将评估科尔纤维密度作为一个 成像生物标志物来预测肿瘤侵袭性和淋巴结转移。科尔纤维将 通过光学SHG显微镜检测，光学SHG显微镜是一种高灵敏度和非侵入性的内在成像技术， 来自科尔纤维信号。我们观察到淋巴结阳性（LN+）的科尔纤维密度增加。 与使用来自乳腺癌的离体原发性肿瘤标本的淋巴结阴性（LN-）患者相比 患者科尔纤维的分子光学显微内镜SHG成像，直接显示ECM 完整性和结构，可以作为预测转移的替代标志物。我们将检验这一假设 三个目标。在目标1中，我们将定量表征原发性肝癌中SHG检测到的科尔纤维特征， 乳腺癌患者的肿瘤标本，并评估定量科尔纤维参数的纤维 “距离”和“纤维体积”预测淋巴结状态。在目标2中，我们将开发、测试和优化 超紧凑型光纤SHG内窥镜检测科尔纤维。在目标3中，我们将执行第一个 在标准超声引导乳腺检查期间使用该光纤SHG显微内镜的临床可行性研究 活检程序，从而获得SHG科尔图像，可以帮助预测淋巴结转移。在 目的1和3，我们还将科尔纤维密度与标准乳腺癌预后标志物相关联， 确定CoH纤维密度唯一选择淋巴结转移风险患者人群的能力 淋巴结受累和潜在的全身性疾病。从更广泛的角度来看，正在建造的设备还将 可用于未来的应用，例如检测对基质消耗疗法的反应， 在桥浆瘤中的递送，以及预测药物递送。

In Research Component 4, we will develop and clinically test endomicroscopic Collagen I (Coll) fiber imaging with an innovative ultra-compact fiber-optic biopsy needle-compatible endomicroscope that is optimized for second harmonic generation (SHG) microscopic detection of Coll fibers. We will integrate this endomicroscope with a routinely used clinical 14-gauge biopsy needle for SHG Coll imaging in preclinical and clinical studies. Coll fibers will be detected in primary breast tumors at the time of biopsy to predict lymph node involvement in breast cancer patients. The assessment of lymph node involvement, which is the most important predictor of prognosis, early on in the clinical management of breast cancer patients, would be of significant advantage in selecting treatment options at the early time of the initial breast biopsy. Coll fibers are an important component of the extracellular matrix (ECM) in breast tumors, and increased stromal Coll fibers facilitate breast tumor formation, invasion, and metastasis. High Coll fiber density in primary tumors promotes metastasis as cancer cells migrate along dense CoH fiber avenues within metastatic primary tumors to form metastatic nodules. We will therefore evaluate Coll fiber density as an imaging biomarker to predict tumor aggressiveness and lymph node metastasis. Coll fibers will be detected by optical SHG microscopy, a highly sensitive and noninvasive technique for imaging intrinsic signal from Coll fibers. We observed an increased Coll fiber density in lymph node positive (LN+) compared to lymph node negative (LN-) patients using ex vivo primary tumor specimens from breast cancer patients. Molecular optical endomicroscopic SHG imaging of Coll fibers, which directly reveals ECM integrity and structure, may serve as a surrogate marker to predict metastasis. We will test this hypothesis in three Aims. In Aim 1, we will quantitatively characterize SHG-detected Coll fiber signatures in primary tumor specimens from breast cancer patients and evaluate if the quantitative Coll fiber parameters 'fiber distance' and 'fiber volume' predict lymph node status. In Aim 2, we will develop, test, and optimize an ultra-compact fiber-optic SHG endomicroscope to detect Coll fibers. In Aim 3, we will perform the first clinical feasibility study with this fiber-optic SHG endomicroscope during standard ultrasound-guided breast biopsy procedures and thereby obtain SHG Coll images that can help predict lymph node metastasis. In Aims 1 and 3, we will also correlate Coll fiber density with standard breast cancer prognostic markers to determine the ability of CoH fiber density to uniquely select a population of patients that are at risk for lymph node involvement and potentially systemic disease. On a wider perspective, the device being built will also be useful for future applications such as detecting response to stromal depletion therapies that improve drug delivery in desmoplasmic tumors, and predicting drug delivery.