Early detection of breast cancer using markers in nipple aspirate fluid, blood, and tumor material from women with breast cancer compared to healthy women.

We previously showed that we can successfully obtain nipple fluid by aspiration and assess microRNA expression levels isolated from this fluid. For further validation of the clinical value of this potential novel screening method, it is crucial to determine how well microRNA expression patterns discriminate NAF derived from cancerous and healthy breasts. Therefore, we will preoperatively aspirate nipple fluid from women having just been diagnosed with or suspected of breast cancer, and compare microRNA expression patterns in nipple fluid derived from cancerous breasts, contralateral unaffected breasts, and healthy controls. In addition we will determine microRNA expression levels in the tumor and blood of the breast cancer patient. Our ultimate goal is, through monitoring of biomarkers in nipple fluid, to develop a reliable method that on one hand allows postponing or avoiding mutilating prophylactic breast surgery as much as possible (reducing morbidity) while on the other hand determining the right time to intervene, thereby preventing development of invasive breast cancer (reducing mortality).

Circulating miRNAs can be easily measured in body fluids which, in case of nipple fluid, could offer a promising minimally invasive method to complement current breast cancer screening methods, especially for high-risk women. In this study we aim to assess the microRNA expression levels in nipple aspiration fluids of women with recently diagnosed breast cancer where we will compare the results to those obtained in healthy controls (which is already available at our department; study NL41845.041.12). The ultimate goal is to more accurately predict the onset of breast cancer.

Multicenter cross-sectional observational study

In this monocenter, cross-sectional study, nipple aspiration will be performed preoperatively in women that are recently diagnosed with, or are suspected to have breast cancer.



Nipple fluid collection:

Nipple fluid will be collected by qualified and trained study UMC Utrecht personnel at the UMC Utrecht.

This will be done before surgery in patients with biopsy proven invasive breast cancer, or in BIRADS 4/5 patients by mammography before taking the core needle biopsy for histological diagnosis.

In short:

1) The subjects are given one spray (4 IE) of oxytocin in both nostrils;

2) The nipple is gently wiped with Nuprep Skin Prep Gel in order to remove keratin plugs and subsequently cleansed with ethanol (see separate attachment);

3) Nipple fluid will be obtained by means of a manual device: A suction cup (aspirator) is placed over the nipple. Repeated gentle suction by a syringe (10-20 cc) draws fluid from inside the duct to the nipple surface. Fluid droplets are collected by capillary tubes. The entire procedure is repeated at the other breast.



Drawing blood

Three vials of blood will be drawn. This is not a mandatory part of the study and the patients will be asked for separate consent.

An earlier study showed that the burden associated with nipple fluid aspiration is minimal and the procedure is well tolerated. To further minimalize any discomfort, nipple fluid will as much as possible be collected at the time of regular visits to the hospital or outpatient clinic. The risks of the procedure are negligible.

However, a few steps within our protocol could potentially lead to side effects, consisting of:



Oxytocin nasal spray: painful contraction of the uterus (sometimes; 0.1-1%); headache, nausea, vomiting, allergic skin reaction (rarely; 0.01-0.1%).



Nipple numbing cream: redness, skin discoloration, skin thickening at site of application (often; 1-10%); warm, sensitive, burning, itching feeling of the skin (sometimes; 0.1-1%)



Nipple fluid aspiration: sensitive or painful breasts, skin irritation, redness of bruising (mild/seldom)

Blood collection: bruising; lightheadedness; fainting, infection at the site where the needle was inserted into the skin