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下的I型胶原(COIL)纤维 使用创新的超紧凑型光纤活检针兼容内窥镜进行成像， 针对COL光纤的二次谐波产生(SHG)显微检测进行了优化。我们将整合这一点 临床上常用的14号活检针用于临床前期SHG Coll成像的内窥镜检查 和临床研究。在活检时会在原发乳腺肿瘤中检测到COL纤维以预测 乳腺癌患者的淋巴转移情况。对淋巴结受累的评估，这是 最重要的预后预测指标，在乳腺癌患者的临床治疗早期， 在最初乳房活检的早期选择治疗方案将是显著的优势。 COIL纤维是乳腺肿瘤细胞外基质(ECM)的重要组成部分，并增加 间质胶原纤维促进乳腺肿瘤的形成、侵袭和转移。高COL纤维密度 原发肿瘤促进转移，因为癌细胞沿着致密的COH纤维途径在 转移性原发肿瘤形成转移性结节。因此，我们将评估COIL纤维密度作为 预测肿瘤侵袭性和淋巴结转移的影像生物标记物。Coll纤维将是 用光学倍频显微镜检测，一种高灵敏度和非侵入性的本征成像技术 来自Coll光纤的信号。我们观察到淋巴结阳性(LN+)的Coll纤维密度增加。 用体外乳腺癌原发肿瘤标本与淋巴结阴性(LN-)患者进行比较 病人。COL纤维的分子光学内窥镜倍频成像，直接显示ECM 完整性和结构可作为预测转移的替代标记物。我们将检验这一假设 有三个目标。在目标1中，我们将定量表征初级的SHG检测到的COIL纤维特征 乳腺癌患者的肿瘤标本及其定量胶原纤维参数的评价 “距离”和“纤维体积”可以预测淋巴结状态。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COLL纤维。在目标3中，我们将执行第一个 该光纤SHG内窥镜在标准超声引导乳房手术中的临床可行性研究 进行活组织检查，从而获得有助于预测淋巴结转移的SHG Coll图像。在……里面 目标1和3，我们还将把Coll纤维密度与标准乳腺癌预后标记物关联起来 确定COH纤维密度唯一地选择有淋巴风险的患者的能力 结节受累和潜在的系统性疾病。从更广泛的角度来看，正在制造的设备还将 对未来的应用很有用，例如检测对改善药物的基质耗竭疗法的反应 结缔组织瘤中的给药，并预测药物给药。

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.