NEW YORK – Researchers from the University of California, San Diego have developed a microfluidic-based adhesion assay that they hope can be used to risk stratify breast cancer patients.

纽约——加州大学圣地亚哥分校的研究人员开发了一种基于微流控的粘附测定法，他们希望可以用于对乳腺癌患者进行风险分层。

Building on nearly

建立在将近

a decade of work

十年的工作

studying the biology of cell adhesion, the researchers decided to use the assay to determine whether there are differences in adhesion between non-malignant, non-metastatic cell lines and malignant and metastatic cell lines, said Adam Engler, a professor of bioengineering at UCSD and one of the test's developers..

研究细胞粘附生物学的研究人员决定使用该检测方法来确定非恶性、非转移性细胞系与恶性及转移性细胞系之间是否存在粘附差异，加州大学圣地亚哥分校生物工程教授、该测试的开发者之一亚当·恩格勒说道。

The research team discovered that non-malignant cells 'stuck like glue' to their environment and had 'very little migration capacity' and 'strong adhesion to their extracellular environment,' while the metastatic cells did not, Engler said. The researchers' findings were first published in 2017, and a .

恩格勒说，研究团队发现非恶性细胞“像胶水一样黏附”在周围环境中，“迁移能力很小”，而且“与细胞外环境有很强的黏附性”，而转移性细胞则没有。研究人员的这一发现最早发表于2017年。

new paper

新论文

released earlier this month in

本月初发布

Cell Reports

细胞报告

builds on the earlier work.

建立在早期工作的基础上。

In the most recent work, Engler and his colleagues looked at the adhesion of cells in mice and humans and found that in mice, weakly adherent cells form secondary metastases at a 'much higher rate' than the strongly adherent cells, Engler said. In addition to the cells' behavior in mice, the researchers also looked at how cells behaved in 16 human patients with normal breast tissue, ductal carcinoma in situ (DCIS), and invasive lobular or ductal carcinoma..

在最近的研究中，恩格勒及其同事观察了小鼠和人类细胞的粘附情况，发现与粘附性强的细胞相比，粘附性弱的细胞在小鼠体内形成继发性转移的“频率高得多”，恩格勒说道。研究人员除了观察细胞在小鼠中的行为外，还研究了 16 名人类患者细胞的行为，这些患者包括正常乳腺组织、导管原位癌 (DCIS) 以及浸润性小叶癌或导管癌。

In the mice, adhesion retrospectively predicted metastatic disease with 100 percent specificity, 85 percent sensitivity, and an area under the curve of .94, according to the

在小鼠中，黏附性回顾性预测转移性疾病具有 100% 的特异性、85% 的敏感性以及 0.94 的曲线下面积，根据

Cell Reports

细胞报告

paper.

纸。

In human samples, 'the overall single-cell population recovered from healthy breast tissue was most strongly adherent, cells from DCIS had intermediate average population adhesion strength, and cells obtained from IDC and ILC tumors were the most weakly adherent across patients,' the researchers wrote.

研究人员写道：“在人体样本中，从健康乳腺组织中提取的总体单细胞群的黏附性最强，来自导管原位癌（DCIS）的细胞具有中等平均群体黏附强度，而从浸润性导管癌（IDC）和浸润性小叶癌（ILC）肿瘤中提取的细胞在患者间表现为最弱的黏附性。”

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The team's work leverages a microfluidic device with a variable width chamber, or a divergent-wall parallel-plate flow chamber, to analyze cells from resected tumors, Engler said. The device has a wide inlet that tapers inward to the chamber that is coated with adhesive proteins found in breast tissue where the tumor cells are placed and pictures are taken.

恩格勒说，该团队的工作利用了一种具有可变宽度腔室的微流体装置，或者说是具有发散壁的平行板流动腔室，来分析从切除的肿瘤中提取的细胞。该装置有一个宽入口，向内逐渐变细到涂有乳腺组织中发现的粘附蛋白的腔室，肿瘤细胞被放置在其中并进行拍照。

The team then pumps fluid through the device, causing the cells to detach from the adhesive proteins and allowing the researchers to see whether the cells were classifed as weakly adherent or strongly adherent. .

团队随后将液体泵入装置，使细胞脱离粘附蛋白，研究人员以此观察细胞是属于弱粘附还是强粘附。

The researchers found that there is a correlation between patients that don't have disease and have very sticky cells and patients who have 'invasive disease' and have very weakly adherent cells, Engler said.

研究人员发现，没有疾病且细胞粘性很强的患者与患有“侵入性疾病”且细胞粘附性很弱的患者之间存在相关性，恩格勒说道。

The microfluidic device the researchers used to determine the adherence of the cells has been patented, and Engler and his colleagues have created a to-be-named startup to license the patent and commercialize the device.

研究人员用来确定细胞粘附力的微流体装置已获得专利，恩格勒和他的同事创建了一家尚未命名的初创公司来授权该专利并商业化该设备。

The researchers hope to use the assay for patients with very low-grade tumors whose cells are proliferating but haven't started to disseminate or migrate, because those patients' outcomes can be quite variable, Engler said. 'It is hard, given current histological markers … to assess whether a patient with DCIS will eventually develop metastatic disease or not,' but hopefully a device like this will give clinicians an additional data point alongside histological analysis to help determine how to best treat their patients, he added..

研究人员希望对那些肿瘤级别非常低的患者使用该检测方法，这些患者的细胞正在增殖，但尚未开始扩散或迁移，因为这些患者的结果可能会有很大差异，恩格勒说道。“根据目前的组织学标记物…很难评估DCIS患者最终是否会发展为转移性疾病，”他补充道，但希望像这样的设备能够为临床医生提供一个与组织学分析相结合的额外数据点，以帮助确定如何最好地治疗他们的患者。

According to Engler, the test could be used as a screening tool using core needle biopsy samples similar to those that are taken for histological analyses, although clinical use of their technology on human patients could be a ways off.

根据恩格勒的说法，这项测试可以作为一种筛查工具，使用与组织学分析类似的核心针刺活检样本，尽管将其技术应用于人类患者的临床使用可能还有段路要走。

The team is currently enrolling more DCIS patients to show stronger correlation between patients who have a higher proportion of weakly adherent cells and those who develop metastatic disease. In the

该团队目前正在招募更多的导管原位癌患者，以展示弱粘附细胞比例较高的患者与发生转移性疾病的患者之间更强的相关性。在

Cell Reports

细胞报告

paper, the researchers found that DCIS patients have a wide range of adhesion types, so the team plans to follow those patients over the next five years as their disease progresses to determine whether the assay was correct and weakly adherent cells were indicative of metastasis and poorer outcomes.

在论文中，研究人员发现导管原位癌患者有多种不同类型的粘附，因此团队计划在接下来的五年里对这些患者进行跟踪观察，随着他们疾病的进展确定该检测是否正确，弱粘附细胞是否预示着转移和更差的结果。

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'When an oncologist orders our test in the future, the outcome of that test could help better decide what the patient's standard of care should be,' Engler said. If a patient has DCIS and weakly adherent cells, a clinician may decide to treat the tumor more aggressively.

“当肿瘤科医生在未来订购我们的测试时，该测试的结果可能有助于更好地决定患者的护理标准应该是什么，”恩格勒说。如果患者患有导管原位癌且细胞粘附性弱，临床医生可能会决定更积极地治疗肿瘤。

For now, the utility of the device for detecting cancer in humans is still largely unproven and additional validation is needed to determine cutoff ranges for what counts as weak adherence versus strong adherence, but the researchers are 'hopeful' that additional validation will show clinical utility in humans, Engler said.

目前，该设备在检测人类癌症方面的实用性仍未得到充分验证，还需要进一步验证来确定弱粘附与强粘附的临界范围，但研究人员“希望”进一步验证能显示出其在人体中的临床实用性，恩格勒说道。

The startup plans to fund a large clinical trial with hundreds of DCIS patients to more accurately determine the cutoffs and stratify patients by their risk. .

这家初创公司计划为涉及数百名导管原位癌患者的大型临床试验提供资金，以更准确地确定临界值，并根据患者的风险进行分层。

The clinical version of the assay would report a number between zero and 1,000 as a result, with the closer a patient is to 1,000 indicating stickier cells. But if a patient has a value near zero, it means the cells aren't adhering to anything and thus won't migrate. There is a 'sweet spot' where the most aggressive tumors are located, and the validation trial will help determine the boundaries of that sweet spot, Engler said..

该检测的临床版本将报告一个介于0到1000之间的数值作为结果，数值越接近1000表示细胞越粘。但如果患者的数值接近零，则意味着细胞没有粘附在任何东西上，因此不会迁移。恩格勒表示，最恶性的肿瘤位于一个“最佳点”，验证试验将有助于确定这个最佳点的边界。

As Engler and his colleagues move toward licensing the patent and commercializing the device, there is still work to be done outside of validation trials. He noted that the device will need to be repackaged into a 'more user-friendly interface' so it is easier to use and more automated, although all of the computer coding and microfluidics work for the device has been done.

恩格勒及其同事在走向专利授权和设备商业化的过程中，除了验证试验之外，还有其他工作需要完成。他指出，该设备需要重新包装成“更用户友好的界面”，以便更容易使用且更加自动化，尽管设备的所有计算机编码和微流体工作已经完成。

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The device could be applied to more than just breast cancer, Engler added. One of the driving forces behind the original research was the search for a 'universal marker' of cancer that was physical in nature rather than biological. The researchers wondered whether 'physical markers are going to be universal because the behaviors are universal' in metastatic cancers, he said.

恩格勒补充说，该设备不仅可以应用于乳腺癌。推动最初研究的其中一个动力是寻找一种物理性质而非生物性质的“癌症通用标记”。他说，研究人员想知道“转移性癌症中的物理标记是否将会是通用的，因为其行为是通用的”。

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'All of those solid tumors start off as a mass in the host tissue, and then the cells have to disseminate somehow,' he noted. 'Physical principles could be applied either to the growth of those tumors or to the spread of those tumors.'

“所有这些实体瘤最初都是在宿主组织中形成的肿块，然后这些细胞必须以某种方式扩散，”他指出。“物理原理可以应用于这些肿瘤的生长或扩散。”

'As cells migrate out of the tumor, the cells that are leaving the tumor versus the ones that are staying in the tumor have to have some physical difference,' Engler added.

“当细胞从肿瘤中迁移出来时，离开肿瘤的细胞与留在肿瘤中的细胞必须存在某种物理差异，”恩格勒补充道。

The researchers looked at lung and prostate cancer cell lines that were more aggressive to see if they had the same signature as aggressive breast cancer cell lines and found that they did.

研究人员观察了更具侵袭性的肺癌和前列腺癌细胞系，以查看它们是否与侵袭性乳腺癌细胞系具有相同的特征，并发现它们确实如此。

'The concept of cell adhesion being really important in the metastatic process at some point' is probably correct 'across all solid tumor types,' Engler said. One of the graduate students within Engler's laboratory is currently investigating pancreatic cancer cells to determine how universal the concept of cell adhesion and metastases is, he added..

“细胞粘附在癌症转移过程中非常重要”这一概念在所有实体肿瘤类型中可能是正确的，恩格勒说道。他补充道，其实验室的一名研究生目前正在研究胰腺癌细胞，以确定细胞粘附和癌症转移的概念具有多大的普遍性。