# Rapid Protease Profiling with a Multiplex Electronic Method for Detection of Metastatic Triple-Negative Breast Cancer

> **NIH NIH R01** · KANSAS STATE UNIVERSITY · 2020 · $278,547

## Abstract

Early diagnosis via cost-effective screening of high risk subpopulation followed by effective
treatments is the key for saving many lives of cancer patients. Proteases overexpressed in
cancer cells and secreted into circulation have been used as drug targets for the development
of protease inhibitors as well as biomarkers for diagnosis and therapeutic monitoring. In this
project, a multi-discipline team intends to develop a nanoelectronic chip for rapid profiling
activity of six proteases that are overexpressed in metastatic triple-negative breast cancer
(TNBC) patients and use this information to identify signatures in breast cancer progression and
therapeutic responsiveness. The team includes (1) Dr. Jun Li (PI, nanotechnology and
biosensor, Kansas State University); (2) Dr. Duy H. Hua (synthetic medicinal chemistry, Kansas
State University); (3) Dr. Meyya Meyyappan (nanotechnology and nanodevices, NASA Ames
Research Center); and (4) Dr. Priyanka Sharma (cancer clinical research, the University of
Kansas Medical Center).
This project focuses on developing the PI’s previously demonstrated electronic device based on
carbon nanofiber nanoelectrode arrays for simultaneous detection of the activity of six
overexpressed proteases in TNBC, including cathepsin B, MMP-2, MMP-9, ADAM-10, ADAM-
17 and uPA. Hexapeptide substrates highly specific to the aforementioned proteases are
covalently attached to the tips of embedded carbon nanofibers in a multiplexed electronic chip.
The distal end of the hexapeptides is attached with an electrochemical reporter. Introducing the
TNBC patient samples containing these proteases results in cleavage of corresponding
hexapeptides, thereby releasing the electrochemical reporter and leading to an exponential
decrease of the electrochemical signal. The protease activity can be derived from the decay
time constant of the kinetic curves.
This extremely sensitive biosensor technology will be fabricated into independently addressed
3x3 arrays and packaged in disposable fluidic cartridges. Using 18 hexapeptide substrates (3
for each protease) attached to different nanoelectrode arrays in two 3x3 electronic chips,
reliable proteolytic activity profile of the aforementioned six proteases in TNBC can be quickly
measured, which cannot be done with current technologies. This technique should greatly
accelerate the speed of protease profiling in complex samples and facilitate detection of
invasive TNBC from other breast cancers in its early stage. The change of the protease activity
profile will be monitored and correlate it with the longitudinal cancer treatment responses.

## Key facts

- **NIH application ID:** 9971473
- **Project number:** 5R01CA217657-04
- **Recipient organization:** KANSAS STATE UNIVERSITY
- **Principal Investigator:** JUN LI
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $278,547
- **Award type:** 5
- **Project period:** 2017-08-01 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9971473

## Citation

> US National Institutes of Health, RePORTER application 9971473, Rapid Protease Profiling with a Multiplex Electronic Method for Detection of Metastatic Triple-Negative Breast Cancer (5R01CA217657-04). Retrieved via AI Analytics 2026-05-29 from https://api.ai-analytics.org/grant/nih/9971473. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*